1 mg/kg: Six-week-old B-NDG female mice (Biocytogen, China) were injected subcutaneously with 5 × 106 MV4-11 cells. Tumor growth was monitored, and mice were assigned to four treatment groups when tumor volumes reached approximately 100 mm3. Each group received two intravenous tail vein injections of one of the following treatments: vehicle or LA-057-MMAE (1, 3, and 6 mg/kg) on day 0 and 7. Tumor volume (V) = (LxW2)/2, where L is the longest dimension of the tumor and W is the corresponding tumor width. Mice were monitored for survival until they either succumbed to the disease or were euthanized when tumor volumes exceeded 1500 mm3.

3 mg/kg: Six-week-old B-NDG female mice (Biocytogen, China) were injected subcutaneously with 5 × 106 MV4-11 cells. Tumor growth was monitored, and mice were assigned to four treatment groups when tumor volumes reached approximately 100 mm3. Each group received two intravenous tail vein injections of one of the following treatments: vehicle or LA-057-MMAE (1, 3, and 6 mg/kg) on day 0 and 7. Tumor volume (V) = (LxW2)/2, where L is the longest dimension of the tumor and W is the corresponding tumor width. Mice were monitored for survival until they either succumbed to the disease or were euthanized when tumor volumes exceeded 1500 mm3.

6 mg/kg: Six-week-old B-NDG female mice (Biocytogen, China) were injected subcutaneously with 5 × 106 MV4-11 cells. Tumor growth was monitored, and mice were assigned to four treatment groups when tumor volumes reached approximately 100 mm3. Each group received two intravenous tail vein injections of one of the following treatments: vehicle or LA-057-MMAE (1, 3, and 6 mg/kg) on day 0 and 7. Tumor volume (V) = (LxW2)/2, where L is the longest dimension of the tumor and W is the corresponding tumor width. Mice were monitored for survival until they either succumbed to the disease or were euthanized when tumor volumes exceeded 1500 mm3.

LA-057-MMAE or isotype control-MMAE was serially diluted in cell culture media and added to AML cells (U937, MV-4-11, and HL60) at a density of 1 × 104 cells per well. The cells were incubated for different durations: U937 cells for 2 days, HL-60 cells for 6 days, and MV4-11 cells for 3 days. For AML patient PBMCs, 3000 cells per well were incubated with serially diluted ADC in StemSpan Leukemic Cell Culture kit (STEMCELL Technologies, USA) for 7 days. Cell viability was measured (Promega) and calculated the growth inhibition of each well by a percentage of cell viability relative to blank wells. The assays were conducted following the guidelines established by Junx Bio for the use of PBMCs from AML patients.

LA-057-MMAE or isotype control-MMAE was serially diluted in cell culture media and added to AML cells (U937, MV-4-11, and HL60) at a density of 1 × 104 cells per well. The cells were incubated for different durations: U937 cells for 2 days, HL-60 cells for 6 days, and MV4-11 cells for 3 days. For AML patient PBMCs, 3000 cells per well were incubated with serially diluted ADC in StemSpan Leukemic Cell Culture kit (STEMCELL Technologies, USA) for 7 days. Cell viability was measured (Promega) and calculated the growth inhibition of each well by a percentage of cell viability relative to blank wells. The assays were conducted following the guidelines established by Junx Bio for the use of PBMCs from AML patients.

LA-057-MMAE or isotype control-MMAE was serially diluted in cell culture media and added to AML cells (U937, MV-4-11, and HL60) at a density of 1 × 104 cells per well. The cells were incubated for different durations: U937 cells for 2 days, HL-60 cells for 6 days, and MV4-11 cells for 3 days. For AML patient PBMCs, 3000 cells per well were incubated with serially diluted ADC in StemSpan Leukemic Cell Culture kit (STEMCELL Technologies, USA) for 7 days. Cell viability was measured (Promega) and calculated the growth inhibition of each well by a percentage of cell viability relative to blank wells. The assays were conducted following the guidelines established by Junx Bio for the use of PBMCs from AML patients.