Fig. 4

Targeting ABCC3 in KPC primary cell line reduces cell proliferation through STAT3 and HIF1α dysregulation. Primary cell lines were established from the pancreatic tumours of KPC mice as described in the Methods section. KPC cells were treated with indicated doses of MCI-715 and number of cells was assessed after 72 h. For the assessment of anchorage-independent growth, cells were plated on soft agar, treated with indicated concentrations of MCI-715 and number of colonies was counted after 4 weeks. a Data presents the effects of MCI-715 dose response treatment in established KPC primary cell lines on anchorage dependent and independent cell growth. Blue: I^st established cell line, Green: 2nd established cell line. Data is expressed as fold change of cells treated with vehicle (DMSO). The results are presented as mean ± SEM of 5 (blue) and 3 (green) independent experiments, One-was ANOVA was performed for statistical analysis, *p < 0.5, **p < 0.01, ***p < 0.001, ****p < 0.0001; Representative Western blot images showing the effects of knockdown of ABCC3 with two specific siRNAs b and treatment with 10 μM MCI-715 c on the expression of pSTAT3 Y705 and HIF1α in the KPC primary cell line. Cells were collected 48 h post transfection/treatment. Quantitative analysis is presented as mean ± SEM of 3 (pSTAT3 Y705) and 4 (HIF1α) independent experiments and expressed as fold change of expression in control sample in reference to loading control; unpaired Student’s t-test was performed for statistical analysis, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001