Gap Junctions Are Involved in the Rescue of CFTR-Dependent Chloride Efflux by Amniotic Mesenchymal Stem Cells in Coculture with Cystic Fibrosis CFBE41o- Cells

Figure 4

Effect of Cx43 siRNA on CFTR protein localisation, expression, and function. (a) Confocal microscopy of cocultures after 6 days from the transfection with either siRNA against CX43 or scrambled siRNA. Cocultures were stained with an anti-CFTR primary antibody followed by an Alexa Fluor 488-conjugated secondary antibody (green) and DAPI (blue). XZ images are shown. Original magnification: 60x. Scale bar: 10 μm. (b) Representative Western blot of a typical experiment analysed using anti-human CFTR C-terminus antibody (dilution 1 : 500), anti-Cx43 (dilution 1 : 500), anti β-tubulin (dilution 1 : 1000). β-Tubulin was used to normalise protein loading. The histogram summarises the relative change in the ratio between band C and band B in the various conditions compared to the CFTR ratio in the 16HBE set as 1. Results represent means ± SEM of three experiments. . (c) CFTR-dependent chloride efflux analysed under the same experimental conditions. Data are shown as mean ± SD of three experiments. .

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