Fig 3. KCNN4 hit confirmation.
(A) OCR of Mia PaCa-2 cells transfected with individual siRNAs against KCNN4, 20 000 cells/well were re-seeded the night before the assay to avoid cell number-dependent changes. n = 6; (B) Upper panel: KCNN4 TaqMan gene expression assay. KCNN4 mRNA levels were normalized to HPRT. KCa3.1 Lower panel: Western Blot analysis of KCNN4 protein expression, HSP90 was used as loading control. Arrow indicates the KCNN4 protein band. (C) Oxygen consumption measurements of Mia PaCa-2, (D) Panc-1, (E) Capan-1 and (F) BxPC-3 cells treated with rac-16 (KCNN4 inhibitor) at different concentrations as indicated, 100 nM NS309 (KCNN4 activator) and high concentrations of extracellular KCl (60 mM), n = 5. Data are represented as the mean ± SD. ANOVA test with Bonferroni post-hoc was performed to compare NT siRNA and siRNA targeting KCNN4 mRNA; ns, not significant, p>0.05, *, p-value is 0.01 to 0.05; **, p-value is < 0.01.