Figure S4.
Effect of FIP200 or WIPI2 deficiency on peroxisome clustering and number upon expression of GFP or OPTN-GFP. HEK-293 cells lacking FIP200 (ΔFIP200, 293) and HEK-293A cells lacking WIPI2 (ΔWIPI2, 293A), along with their respective parental control cell lines (CT), were transfected with plasmids encoding either GFP or OPTN-GFP (OPTN-G). After 2 days, cells were processed for IF microscopy using antibodies against PEX14. Nuclei were counterstained with DAPI. (A) Representative images are shown. Scale bars: 10 µm (insets: 50 µm). (B) Quantification of relative peroxisome number. Each dot represents a single cell. Data are expressed as mean ± SD. To minimize imaging bias, all images were acquired at the nuclear focal plane to ensure consistency across conditions. Statistical comparisons are indicated (ns, nonsignificant; *, P < 0.05; **, P < 0.01).

Effect of FIP200 or WIPI2 deficiency on peroxisome clustering and number upon expression of GFP or OPTN-GFP. HEK-293 cells lacking FIP200 (ΔFIP200, 293) and HEK-293A cells lacking WIPI2 (ΔWIPI2, 293A), along with their respective parental control cell lines (CT), were transfected with plasmids encoding either GFP or OPTN-GFP (OPTN-G). After 2 days, cells were processed for IF microscopy using antibodies against PEX14. Nuclei were counterstained with DAPI. (A) Representative images are shown. Scale bars: 10 µm (insets: 50 µm). (B) Quantification of relative peroxisome number. Each dot represents a single cell. Data are expressed as mean ± SD. To minimize imaging bias, all images were acquired at the nuclear focal plane to ensure consistency across conditions. Statistical comparisons are indicated (ns, nonsignificant; *, P < 0.05; **, P < 0.01).

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