Fig. 3
From: APOL1 plasma membrane pools resist rapid protein degradation
Rapid APOL1 degradation is isoform-independent. HEK293T cells allowing the conditional expression of C-terminal GFP-tagged APOL1 splice variants (G0) with and without MG132 treatment. (A) Scheme: Membrane orientation of APOL1 isoforms. Left: APOL1 isoforms vA, and vB1 (vC) with N- and C-termini facing the ER lumen. Right: APOL1 isoform vB3 and APOL2 with both termini on the cytoplasmic side. (B) Composed histograms of FC analyses with APOL1 G0-GFP isoforms vA, vB1, vB3 and vC expressing cells without further treatment (+ Dox), or in combination with MG132 or cycloheximide (CHX) respectively. The histograms show representative FC analyses of at least three independent experiments (N ≥ 3); y-axis: cell count; x-axis: GFP-fluorescence. (C,D) Graphs: FC analyses of the MG132 (C) and CHX (D) treatments shown as MFI values. -Dox: non-induced cells; +Dox: induced for 24 h without inhibitor and with MG132 or CHX treatment for indicated time periods (N ≥ 3); y-axis: cell count; x-axis: GFP-fluorescence. ns: not significant, *: p < 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001.
