On the day of administration, HER2 targeted ADC and HER2 were prepared at a 10 fold ratio in standard growth medium. Add the prepared targeted ADC treatment to cells to obtain a final concentration of 3.81e-5nM to 100nM and a final volume of 50uL/well. Each drug concentration is tested in quadruplicate. On the same day, the cell viability of untreated cells at the time of administration was evaluated by adding 25 u L of CellTiter (ProMag catalog number G7573), a reagent for lysing cells and measuring total adenosine triphosphate (ATP) content. Incubate the plate at room temperature for 10 minutes to stabilize the luminescence signal before reading. Use luminous readingThe PHERAstar FSX ELISA reader (BMG Labtech) was used to measure the luminescence signals of all plates. This signal represents the cell viability of untreated cells on the day of administration or day 0.

On the day of administration, HER2 targeted ADC and HER2 were prepared at a 10 fold ratio in standard growth medium. Add the prepared targeted ADC treatment to cells to obtain a final concentration of 3.81e-5nM to 100nM and a final volume of 50uL/well. Each drug concentration is tested in quadruplicate. On the same day, the cell viability of untreated cells at the time of administration was evaluated by adding 25 u L of CellTiter (ProMag catalog number G7573), a reagent for lysing cells and measuring total adenosine triphosphate (ATP) content. Incubate the plate at room temperature for 10 minutes to stabilize the luminescence signal before reading. Use luminous readingThe PHERAstar FSX ELISA reader (BMG Labtech) was used to measure the luminescence signals of all plates. This signal represents the cell viability of untreated cells on the day of administration or day 0.

On the day of administration, HER2 targeted ADC and HER2 were prepared at a 10 fold ratio in standard growth medium. Add the prepared targeted ADC treatment to cells to obtain a final concentration of 3.81e-5nM to 100nM and a final volume of 50uL/well. Each drug concentration is tested in quadruplicate. On the same day, the cell viability of untreated cells at the time of administration was evaluated by adding 25 u L of CellTiter (ProMag catalog number G7573), a reagent for lysing cells and measuring total adenosine triphosphate (ATP) content. Incubate the plate at room temperature for 10 minutes to stabilize the luminescence signal before reading. Use luminous readingThe PHERAstar FSX ELISA reader (BMG Labtech) was used to measure the luminescence signals of all plates. This signal represents the cell viability of untreated cells on the day of administration or day 0.

On the day of administration, HER2 targeted ADC and HER2 were prepared at a 10 fold ratio in standard growth medium. Add the prepared targeted ADC treatment to cells to obtain a final concentration of 3.81e-5nM to 100nM and a final volume of 50uL/well. Each drug concentration is tested in quadruplicate. On the same day, the cell viability of untreated cells at the time of administration was evaluated by adding 25 u L of CellTiter (ProMag catalog number G7573), a reagent for lysing cells and measuring total adenosine triphosphate (ATP) content. Incubate the plate at room temperature for 10 minutes to stabilize the luminescence signal before reading. Use luminous readingThe PHERAstar FSX ELISA reader (BMG Labtech) was used to measure the luminescence signals of all plates. This signal represents the cell viability of untreated cells on the day of administration or day 0.

On the day of administration, HER2 targeted ADC and HER2 were prepared at a 10 fold ratio in standard growth medium. Add the prepared targeted ADC treatment to cells to obtain a final concentration of 3.81e-5nM to 100nM and a final volume of 50uL/well. Each drug concentration is tested in quadruplicate. On the same day, the cell viability of untreated cells at the time of administration was evaluated by adding 25 u L of CellTiter (ProMag catalog number G7573), a reagent for lysing cells and measuring total adenosine triphosphate (ATP) content. Incubate the plate at room temperature for 10 minutes to stabilize the luminescence signal before reading. Use luminous readingThe PHERAstar FSX ELISA reader (BMG Labtech) was used to measure the luminescence signals of all plates. This signal represents the cell viability of untreated cells on the day of administration or day 0.

On the day of administration, HER2 targeted ADC and HER2 were prepared at a 10 fold ratio in standard growth medium. Add the prepared targeted ADC treatment to cells to obtain a final concentration of 3.81e-5nM to 100nM and a final volume of 50uL/well. Each drug concentration is tested in quadruplicate. On the same day, the cell viability of untreated cells at the time of administration was evaluated by adding 25 u L of CellTiter (ProMag catalog number G7573), a reagent for lysing cells and measuring total adenosine triphosphate (ATP) content. Incubate the plate at room temperature for 10 minutes to stabilize the luminescence signal before reading. Use luminous readingThe PHERAstar FSX ELISA reader (BMG Labtech) was used to measure the luminescence signals of all plates. This signal represents the cell viability of untreated cells on the day of administration or day 0.

Cultivate the cell line in a tissue culture incubator at 5% CO2 and 37 °C in the optimal growth medium. Separate cells at least 2 days prior to inoculation for proliferation assays to ensure optimal growth density. On the day of vaccination, cell viability and cell density were determined using a cell counter (Vi Cell XR Cell viability analyzer, Beckman Coulter). Inoculate cells with vitality above 85% into a 384 well TC treated plate (Corning catalog number 3765) with a white transparent bottom. Inoculate cells at a density of 1000 cells per well into 45 u L of standard growth medium. Incubate the plate overnight in a tissue culture incubator at 5% CO2 and 37 °C. On the second day, prepare free atorvastatin payload and targeted ADC at a 10 fold ratio in standard growth medium. Add the prepared treatment substance to the cells to obtain a final concentration of 0.005-100nM and a final volume of 50 u L/well. Each drug concentration is tested in quadruplicate. Incubate the plates in a tissue culture incubator at 5% CO2 and 37 °C for 5 days, and then evaluate cell viability by adding 25 u L of CellTiterGlo (Promega, catalog number G7573) (a reagent for cell lysis and measurement of total adenosine triphosphate (ATP) content). Incubate the plate at room temperature for 10 minutes to stabilize the luminescence signal, and then read using a luminescence reader (EnVision multi label enzyme-linked immunosorbent assay, PerkinElmer). To evaluate the effectiveness of drug treatment, luminescence counts from wells containing untreated cells (100% viability) were used to normalize the treated samples. Apply a variable slope model to fit the nonlinear regression curve to the data in GraphPad PRISM version 7.02 software. Extrapolating IC50 from the obtained curve

Cultivate the cell line in a tissue culture incubator at 5% CO2 and 37 °C in the optimal growth medium. Separate cells at least 2 days prior to inoculation for proliferation assays to ensure optimal growth density. On the day of vaccination, cell viability and cell density were determined using a cell counter (Vi Cell XR Cell viability analyzer, Beckman Coulter). Inoculate cells with vitality above 85% into a 384 well TC treated plate (Corning catalog number 3765) with a white transparent bottom. Inoculate cells at a density of 1000 cells per well into 45 u L of standard growth medium. Incubate the plate overnight in a tissue culture incubator at 5% CO2 and 37 °C. On the second day, prepare free atorvastatin payload and targeted ADC at a 10 fold ratio in standard growth medium. Add the prepared treatment substance to the cells to obtain a final concentration of 0.005-100nM and a final volume of 50 u L/well. Each drug concentration is tested in quadruplicate. Incubate the plates in a tissue culture incubator at 5% CO2 and 37 °C for 5 days, and then evaluate cell viability by adding 25 u L of CellTiterGlo (Promega, catalog number G7573) (a reagent for cell lysis and measurement of total adenosine triphosphate (ATP) content). Incubate the plate at room temperature for 10 minutes to stabilize the luminescence signal, and then read using a luminescence reader (EnVision multi label enzyme-linked immunosorbent assay, PerkinElmer). To evaluate the effectiveness of drug treatment, luminescence counts from wells containing untreated cells (100% viability) were used to normalize the treated samples. Apply a variable slope model to fit the nonlinear regression curve to the data in GraphPad PRISM version 7.02 software. Extrapolating IC50 from the obtained curve

Cultivate the cell line in a tissue culture incubator at 5% CO2 and 37 °C in the optimal growth medium. Separate cells at least 2 days prior to inoculation for proliferation assays to ensure optimal growth density. On the day of vaccination, cell viability and cell density were determined using a cell counter (Vi Cell XR Cell viability analyzer, Beckman Coulter). Inoculate cells with vitality above 85% into a 384 well TC treated plate (Corning catalog number 3765) with a white transparent bottom. Inoculate cells at a density of 1000 cells per well into 45 u L of standard growth medium. Incubate the plate overnight in a tissue culture incubator at 5% CO2 and 37 °C. On the second day, prepare free atorvastatin payload and targeted ADC at a 10 fold ratio in standard growth medium. Add the prepared treatment substance to the cells to obtain a final concentration of 0.005-100nM and a final volume of 50 u L/well. Each drug concentration is tested in quadruplicate. Incubate the plates in a tissue culture incubator at 5% CO2 and 37 °C for 5 days, and then evaluate cell viability by adding 25 u L of CellTiterGlo (Promega, catalog number G7573) (a reagent for cell lysis and measurement of total adenosine triphosphate (ATP) content). Incubate the plate at room temperature for 10 minutes to stabilize the luminescence signal, and then read using a luminescence reader (EnVision multi label enzyme-linked immunosorbent assay, PerkinElmer). To evaluate the effectiveness of drug treatment, luminescence counts from wells containing untreated cells (100% viability) were used to normalize the treated samples. Apply a variable slope model to fit the nonlinear regression curve to the data in GraphPad PRISM version 7.02 software. Extrapolating IC50 from the obtained curve

Cultivate the cell line in a tissue culture incubator at 5% CO2 and 37 °C in the optimal growth medium. Separate cells at least 2 days prior to inoculation for proliferation assays to ensure optimal growth density. On the day of vaccination, cell viability and cell density were determined using a cell counter (Vi Cell XR Cell viability analyzer, Beckman Coulter). Inoculate cells with vitality above 85% into a 384 well TC treated plate (Corning catalog number 3765) with a white transparent bottom. Inoculate cells at a density of 1000 cells per well into 45 u L of standard growth medium. Incubate the plate overnight in a tissue culture incubator at 5% CO2 and 37 °C. On the second day, prepare free atorvastatin payload and targeted ADC at a 10 fold ratio in standard growth medium. Add the prepared treatment substance to the cells to obtain a final concentration of 0.005-100nM and a final volume of 50 u L/well. Each drug concentration is tested in quadruplicate. Incubate the plates in a tissue culture incubator at 5% CO2 and 37 °C for 5 days, and then evaluate cell viability by adding 25 u L of CellTiterGlo (Promega, catalog number G7573) (a reagent for cell lysis and measurement of total adenosine triphosphate (ATP) content). Incubate the plate at room temperature for 10 minutes to stabilize the luminescence signal, and then read using a luminescence reader (EnVision multi label enzyme-linked immunosorbent assay, PerkinElmer). To evaluate the effectiveness of drug treatment, luminescence counts from wells containing untreated cells (100% viability) were used to normalize the treated samples. Apply a variable slope model to fit the nonlinear regression curve to the data in GraphPad PRISM version 7.02 software. Extrapolating IC50 from the obtained curve

Cultivate the cell line in a tissue culture incubator at 5% CO2 and 37 °C in the optimal growth medium. Separate cells at least 2 days prior to inoculation for proliferation assays to ensure optimal growth density. On the day of vaccination, cell viability and cell density were determined using a cell counter (Vi Cell XR Cell viability analyzer, Beckman Coulter). Inoculate cells with vitality above 85% into a 384 well TC treated plate (Corning catalog number 3765) with a white transparent bottom. Inoculate cells at a density of 1000 cells per well into 45 u L of standard growth medium. Incubate the plate overnight in a tissue culture incubator at 5% CO2 and 37 °C. On the second day, prepare free atorvastatin payload and targeted ADC at a 10 fold ratio in standard growth medium. Add the prepared treatment substance to the cells to obtain a final concentration of 0.005-100nM and a final volume of 50 u L/well. Each drug concentration is tested in quadruplicate. Incubate the plates in a tissue culture incubator at 5% CO2 and 37 °C for 5 days, and then evaluate cell viability by adding 25 u L of CellTiterGlo (Promega, catalog number G7573) (a reagent for cell lysis and measurement of total adenosine triphosphate (ATP) content). Incubate the plate at room temperature for 10 minutes to stabilize the luminescence signal, and then read using a luminescence reader (EnVision multi label enzyme-linked immunosorbent assay, PerkinElmer). To evaluate the effectiveness of drug treatment, luminescence counts from wells containing untreated cells (100% viability) were used to normalize the treated samples. Apply a variable slope model to fit the nonlinear regression curve to the data in GraphPad PRISM version 7.02 software. Extrapolating IC50 from the obtained curve