Supplementary Figure 1: Validation of SWELL1 knock-down. Related to Fig. 2.
From: SWELL1 is a regulator of adipocyte size, insulin signalling and glucose homeostasis
(a) qRT-PCR assessment of SWELL1/LRRC8a, and LRRC8b-e mRNA in human adipocyte cell line transduced with Ad-shSWELL1-mCherry (red) compared to Ad-shSCR-mCherry (black; n = 3 culture dishes of adipocytes for each condition). (b) SWELL1 Western blot in human adipocyte cell line transduced with Ad-shSWELL1-mCherry (red) compared to Ad-shSCR-mCherry (black). β-actin serves as loading control. Representative blot from 4 independent experiments. (c) VRAC over time ± hypotonic swelling (210 mOsm) in 3T3-F442A adipocytes transduced with Ad-shSCR-mCherry (black) and Ad-shSWELL1-mCherry (red). (d) Mean peak outward (+100 mV) and peak inward (−100 mV) VRAC current density from (c) in Ad-shSCR (black, n = 5) and Ad-shSWELL1 (red, n = 4) transduced 3T3-F442A adipocytes. (e) VRAC current-voltage relationship after hypotonic swelling (210 mOsm) in human adipocytes transduced with Ad-shSCR-mCherry (black) and Ad-shSWELL1-mCherry (red). (f) Mean peak outward (+100 mV) and peak inward (−100 mV) VRAC current density from (e) in Ad-shSCR (black, n = 10) and Ad-shSWELL1 (red, n = 10) transduced human adipocytes. Uncropped blots for b are shown in Supplementary Fig. 9a–c. Significance between the indicated groups in a,d and f was calculated using a two-tailed Student’s t-test. Exact P-values are listed in Supplementary Table 6. Error bars represent mean ± s.e.m.∗(P < 0.05), ∗∗(P < 0.01).
