Reduction of BAK1 and STAT3 expression may mimic miR-125b–induced phenotypes in a dose-dependent manner. (A) Western blotting (top panel) of BAK1 in 32D cells transduced with shRNA-ctrl and specific anti-BAK1 shRNA (shRNA-BAK1) lentiviruses and proliferation kinetics (bottom panel) of 32D cells expressing shRNA-ctrl (open circle) and shRNA-BAK1 (filled circle) during G-CSF treatment. (B) Western blotting (top panel) of STAT3 in 32D cells transduced with shRNA-ctrl and specific anti-STAT3 shRNA (shRNA-STAT3) lentiviruses with comparable RFP expression and proliferation kinetics (bottom panel) of 32D cells expressing shRNA-ctrl-low RFP⁺ (open circle), shRNA-STAT3-low RFP⁺ (filled circle), shRNA-ctrl-high RFP⁺ (open triangle), and shRNA-STAT3-high RFP⁺ (filled triangle) during G-CSF treatment. (C) Western blotting (top panel) of STAT3 and BAK1 in 32D cells transduced with shRNA-ctrl, shRNA-ctrl/BAK1 and shRNA-STAT3/BAK1 lentiviral vectors. Proliferation kinetics (bottom panel) of 32D/shRNA-ctrl (open circle) and 32D/shRNA-STAT3/BAK1 (filled circle) cells during G-CSF-treatment. All 32D/shRNA-STAT3 derivates were sorted for low RFP expression before lentiviral transduction with shRNA-BAK1. Mean of 4 experiments and representative Western blots are shown, respectively. *P < .05 compared with control.