Fig. 2.

Effect of shRNA-mediated knockdown of the suppressor genes on ΔF508-CFTR channel activity. Average normalized fluorescence intensity of ΔF508-CFTR cells transfected with shRNA for A, FGFR1, B, RIPK4, C, MET, D, SHPK, E, MAP3K13, F, BRAF, G, DUSP22, H, CDK10, I, IPMK, or luciferase (nonsilencing control), and grown at 37 °C. After 48 h ΔF508-CFTR cells were subjected to puromycin selection (3 days) and stimulated with FIG (25 μm Forskolin, 45 μm IBMX, and 50 μm Genistein). Quenching of YFP fluorescence during Cl⁻/I⁻ exchange of 70–300 cells per field was recorded and quantified simultaneously by Cellomics ArrayScan VTI. Multiple shRNA clones per gene were analyzed. One representative shRNA clone is shown (Supplemental Table S2). KD, knockdown efficiency (%). J, Quantitation of rescue (difference in average fluorescence intensity ΔFI_avg) of ΔF508-CFTR at 24 s after adding iodide solution. Data are mean ± S.E. from 2–3 independent experiments (three fields per well, 70–300 cells per field). Inset: Comparison of normalized average fluorescent intensity of ΔF508-CFTR (F508-CFTR) versus WT-CFTR.