Abstract

<p>Immunoblot of CFTR following sip23 treatment of F508del-expressing cells at 37°C (A) and 30°C (B) or WT-expressing cells (C). Histograms show quantification of CFTR band-B and C glycoforms and C/B ratios. Results are shown as a percent of the maximal signal for band-B glycoform and as fold change relative to control (set to 1) for the ratio C/B (mean ± SEM, n≥3). (D) qRT-PCR analysis of CFTR and p23 levels following p23 silencing in WT or F508del-expressing cells. Results represent a ratio of the indicated mRNA to GUS, and shown as the percent of siRNA control (mean ± SEM, n≥3). (E) Iodide efflux analysis of F508del-expressing cells in response to p23 siRNA or 30°C correction. Results are shown as a ratio of the efflux at stimulation (stim) to efflux at pre-stimulation (basal) (mean ± SD, n≥3). (F) Immunoblot of the indicated proteins in the cell lysate (input) or following CFTR IP (right panels) in response to sip23 treatment (CFTR−/− cells were used as a negative control for the CFTR IP). Histogram shows quantification of recovered Hsc/p90 (α and β) and Hsc/p70 by co-IP with CFTR. The data is shown as a ratio of the recovered chaperone to total CFTR and normalized to 1 for the control siRNA (mean ± SD, n = 3, replicated three times). For all data, * indicates <i>p</i><0.05 relative to control, and results were replicated at least once. The underlying data used to make (A–F) in this figure can be found in the supplementary file <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001998#pbio.1001998.s008" target="_blank">Data S1</a>.</p

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