Figure 2.
Figure 2. Overexpressed USP30 reduces peroxisome loss induced by starvation. (A) Immunofluorescence of HeLa cells transfected with USP30-Flag, PMP34-USP30-Flag, or TOM20-USP30-Flag or mock transfected. Cells grown in DMEM or HBSS for 24 h, stained for PMP70 and Flag. (B) Quantification of the peroxisome density in A and E, relative to mock DMEM (n = 3; 30 cells/trial). (C) HeLa lysates expressing USP30-Flag, analyzed by immunoblotting for PEX14 and GAPDH. (D) Quantification of PEX14/GAPDH density ratio in C compared with mock DMEM (n = 6). (E) HeLa cells expressing USP30-Flag C77S or PMP34-USP30-Flag C77S, costained for PMP70. (F and G) HeLa cells expressing PEX26-RG alone (F) or coexpressed with USP30-Flag (G). (H) Quantification of average percent lysosomal peroxisomes (n = 3; 50 cells/trial). Statistical significance (Student’s t test): *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. Bars: 20 µm (A and E); 10 µm (F and G).

Overexpressed USP30 reduces peroxisome loss induced by starvation. (A) Immunofluorescence of HeLa cells transfected with USP30-Flag, PMP34-USP30-Flag, or TOM20-USP30-Flag or mock transfected. Cells grown in DMEM or HBSS for 24 h, stained for PMP70 and Flag. (B) Quantification of the peroxisome density in A and E, relative to mock DMEM (n = 3; 30 cells/trial). (C) HeLa lysates expressing USP30-Flag, analyzed by immunoblotting for PEX14 and GAPDH. (D) Quantification of PEX14/GAPDH density ratio in C compared with mock DMEM (n = 6). (E) HeLa cells expressing USP30-Flag C77S or PMP34-USP30-Flag C77S, costained for PMP70. (F and G) HeLa cells expressing PEX26-RG alone (F) or coexpressed with USP30-Flag (G). (H) Quantification of average percent lysosomal peroxisomes (n = 3; 50 cells/trial). Statistical significance (Student’s t test): *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. Bars: 20 µm (A and E); 10 µm (F and G).

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