Figure 1.
FABP7 deficiency induces cholesterol accumulation in lysosomes. (A–D) SV589 fibroblasts (A and B) and U251 cells (C and D) were transfected with the indicated siRNAs for 48 h, fixed, and stained with filipin (magenta) and the antibody against endogenous LAMP1 (green). Boxed areas are shown at a higher magnification on the right. Scale bars, 10 µm (main), 1 µm (inset). Relative fluorescence intensity of filipin in A and C was quantified by ImageJ and shown in B and D, respectively. Data are presented as median with interquartile range (n = 651 cells from three independent experiments in B, and n = 401 cells from three independent experiments in D). Mann–Whitney U test. Compared with cells transfected with scrambled control siRNA (NC, negative control). (E) Wild-type (WT) and two lines of FABP7 knockout (KO) SV589 fibroblasts (FABP7 KO1 and FABP7 KO2) were fixed and stained with filipin (magenta) and the anti-LAMP1 antibody (green). Boxed areas are shown at a higher magnification on the right. Scale bars, 10 µm (main), 1 µm (inset). (F) Quantification of relative fluorescence intensity of filipin in E. Data are presented as median with interquartile range (n = 356 cells from three independent experiments). Mann–Whitney U test. Compared with WT cells. (G) Relative cholesterol levels of lysosomes purified from WT and FABP7 KO2 cells normalized to protein amounts. Data are presented as mean ± SEM (from four independent experiments). Student’s unpaired two-tailed t test. Compared with WT cells. (H) WT and two lines of FABP7 KO cells (FABP7 KO1 and FABP7 KO2) were fixed, breached with a liquid nitrogen pulse, stained with the purified recombinant His6-EGFP-D4H protein (green) and the anti-LAMP1 antibody (magenta). Boxed areas are shown at a higher magnification on the right. Scale bars, 10 µm (main), 1 µm (inset). (I) Quantification of relative fluorescence intensity of His6-EGFP-D4H protein in H. Data are presented as median with interquartile range (n = 110 cells from three independent experiments). Mann–Whitney U test. Compared with WT cells.

FABP7 deficiency induces cholesterol accumulation in lysosomes. (A–D) SV589 fibroblasts (A and B) and U251 cells (C and D) were transfected with the indicated siRNAs for 48 h, fixed, and stained with filipin (magenta) and the antibody against endogenous LAMP1 (green). Boxed areas are shown at a higher magnification on the right. Scale bars, 10 µm (main), 1 µm (inset). Relative fluorescence intensity of filipin in A and C was quantified by ImageJ and shown in B and D, respectively. Data are presented as median with interquartile range (n = 651 cells from three independent experiments in B, and n = 401 cells from three independent experiments in D). Mann–Whitney U test. Compared with cells transfected with scrambled control siRNA (NC, negative control). (E) Wild-type (WT) and two lines of FABP7 knockout (KO) SV589 fibroblasts (FABP7 KO1 and FABP7 KO2) were fixed and stained with filipin (magenta) and the anti-LAMP1 antibody (green). Boxed areas are shown at a higher magnification on the right. Scale bars, 10 µm (main), 1 µm (inset). (F) Quantification of relative fluorescence intensity of filipin in E. Data are presented as median with interquartile range (n = 356 cells from three independent experiments). Mann–Whitney U test. Compared with WT cells. (G) Relative cholesterol levels of lysosomes purified from WT and FABP7 KO2 cells normalized to protein amounts. Data are presented as mean ± SEM (from four independent experiments). Student’s unpaired two-tailed t test. Compared with WT cells. (H) WT and two lines of FABP7 KO cells (FABP7 KO1 and FABP7 KO2) were fixed, breached with a liquid nitrogen pulse, stained with the purified recombinant His6-EGFP-D4H protein (green) and the anti-LAMP1 antibody (magenta). Boxed areas are shown at a higher magnification on the right. Scale bars, 10 µm (main), 1 µm (inset). (I) Quantification of relative fluorescence intensity of His6-EGFP-D4H protein in H. Data are presented as median with interquartile range (n = 110 cells from three independent experiments). Mann–Whitney U test. Compared with WT cells.

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