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. 2020 Aug 7;2(9):526–537. doi: 10.1096/fba.2020-00012

FIGURE 3.

FIGURE 3

siRNA knockdown demonstrated that pendrin, but not SLC26A3 or SLC26A6, is required for the interleukin‐dependent increase in CFTR current. A, Left, Ussing Chamber recordings from ΔF508/ΔF508 CF HBECs transfected with pendrin (SLC26A4) siRNA and treated with the LIE combination (1 µmol/L Lumacaftor +0.02 µmol/L Ivacaftor +3 µmol/L Elexacaftor) and 10 ng/mL IL‐4 for 48 hours showed that pendrin knockdown prevented the IL‐4 dependent increase in CFTR current (n = 4). Center and right, Ussing Chamber recordings from CF HBECs transfected with either SLC26A3 or SLC26A6 siRNAs showed no significant reduction in the IL‐4 dependent increase in CFTR current (n = 4). B, Taqman RT‐PCR results for SLC26A4 (left), SLC26A3 (center), and SLC26A6 (right) from the same cells as in (A) showed approximately 30% knockdown of target mRNAs. Niflumic acid also reduced SLC26A4 expression, but did not affect SLC26A3 or SLC26A6 (n = 4). C, Western Blot of pendrin in whole cell lysates of epithelia from (A), treated with control or pendrin (SLC26A4) siRNAs, the LIE combination and other agents as indicated. D, Quantitation of the results in C showing concomitant decreases in pendrin protein (normalized to GAPDH) in the SLC26A4 siRNA‐treated epithelia