Journal: bioRxiv
Article Title: ERK plays a conserved dominant role in pancreas cancer cell EMT heterogeneity driven by diverse growth factors and chemotherapies
doi: 10.1101/2025.02.08.637251
Figure Lengend Snippet: HPAF-II cells were transfected with control siRNA or siRNAs targeting ERK1/2 or c-Jun for 24 hr and then treated with 50 ng/mL HGF + 10ng/mL TGFβ for 48 hr. A ) Representative 4i images of pERK and pc-Jun for the indicated conditions. Images representative of n = 3. B) and C) Bar plots displaying fraction of cells in “pc-Jun high” or “pERK high” signaling bins for the indicated conditions. n = 3, one-way ANOVA with Tukey’s multiple comparisons test. D) HPAF-II cells were transfected with control siRNA or siRNAs for the knockdown of ERK1/2 or c-Jun for 24 hr and then treated with 50 ng/mL EGF + 10 ng/mL TGFβ or 50 ng/mL HGF + 10 ng/mL TGFβ for 24 hr. Representative 4i images of pERK and pc-Jun are shown, n = 3. E) and F) For the experiment described in (D), pERK or pc-Jun signal intensity is displayed. n = 3, one-way ANOVA with Tukey’s multiple comparisons test. G) HPAF-II cells were transfected with control siRNA or siRNAs for the knockdown of ERK1/2 or c-Jun for 24 hr and then treated with 50 ng/mL EGF + 10 ng/mL TGFβ or 50 ng/mL HGF + 10 ng/mL TGFβ for 48 hr. Representative 4i images of indicated proteins across treatment and siRNA conditions at 48 hr, n = 3. H) For the experiment described in (G), the mosaic plot displays percentages of cells in each EMT phenotype bin. n = 3, Chi-squared test. I) For the experiment described in (G), representative 4i images of total ERK are shown, n = 3 . J) For the experiment described in (G), the net percent reduction in uncertainty of the EMT phenotype based on pERK, pc-Jun, or both signals is plotted. n=3, one-way ANOVA with Tukey’s multiple comparisons test. For all panels *p < 0.05, and error bars represent standard error of the mean. Scale bars indicate 300 μm.
Article Snippet: Cells were plated on top of the siRNA-lipid complexes 24 hr prior to treatment and fixed for immunofluorescence staining 72 hr post-transfection. siRNA against ERK1/2 (#6560) was purchased from Cell Signal Technology. c-Jun (sc-29223) and control siRNA (sc-37007) were purchased from Santa Cruz Biotechnology.
Techniques: Transfection, Control, Knockdown
