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. 2018 Apr 9;27(11):1927–1940. doi: 10.1093/hmg/ddy101

Figure 4.

Figure 4.

Perinuclear accumulation of cholesterol in SNX14 mutant cells. SNX14 protein in cell lysates from HEK293 clones generated from single cell sorting following CRISPR-Cas9 mediated targeting of the SNX14 gene. (A, B) Clones display either full length SNX14 protein (SNX WT 1–6), no SNX14 protein (SNX14 KO 1–7) or truncated SNX14 protein (SNX14 DEL 1–5). Cells were cultured (C) with or (D) without 23.5 µm U18666A for 24 h. SNX14 mutant cells showed greater accumulation of cholesterol with U18666A treatment. (E) U18666A increased perinuclear distribution of cholesterol. (F) SNX14 mutant clones treated with U18666A displayed an increased perinuclear distribution of cholesterol compared to SNX14WT clones. (C, D) Scale bar = 50 µm. (E, F) The mean radial distribution of filipin signal intensity plotted from nuclear to peripheral regions, N = 6 (SNX14WT clones), N = 5 (SNX14DEL clones), N = 7 (SNX14KO clones), dots = mean, error bars = SD, **P ≤ 0.01, *P ≤ 0.05, n.s. P ≥ 0.05, Student’s t-test.