Fig. 1: HSPB3 is enriched at the nuclear envelope.
From: Small heat-shock protein HSPB3 promotes myogenesis by regulating the lamin B receptor
A Immunofluorescence pictures showing the absence of endogenous HSPB3 (green) in cycling myoblasts (top panel) and its subcellular distribution in 7-day differentiating human myoblasts (lower panel). DAPI staining is shown. Scale bar = 10 µm. Quantification of HSPB3 subcellular distribution in 7-day differentiating human myoblasts is shown. n = 5 independent experiments, ± s.e.m. The total number of cells analyzed: 285. B Immunofluorescence pictures showing that HSPB3 (red) does not colocalize with nuclear HSPB2 (green) foci in differentiating human myoblasts. DAPI staining is shown. Scale bar = 10 µm. C Quantification of HSPB2 and HSPB3 colocalization in differentiating myoblasts. Pearson’s correlation coefficients (PCCs) of images of Alexa Fluor 488-HSPB2 and Alexa Fluor 594-HSPB3 in 7-day differentiating myoblasts cells expressing endogenous HSPB2 and HSPB3 (n = 55 multinucleated myotubes). PCCs of images of Alexa Fluor 488-myc and Alexa Fluor 594-HSPB3 in cycling LHCNM2 cells overexpressing myc-HSPB3 for 24 h, before and after rotating Alexa Fluor 594-HSPB3 image by 90° (n = 47 myoblasts). P < 10-10, +/−s.e.m. D, E 7-day differentiating (D) and cycling (E) human myoblasts were infected with lentiviral particles expressing myc-HSPB3. Immunofluorescence pictures showing colocalization of myc-HSPB3 with endogenous lamin B1 (LMNB1) filaments and at the nuclear envelope. DAPI staining is shown. Scale bar = 10 µm. F Overexpressed GFP-HSPB3 (co-expressed at a 1:8 ratio with myc-HSPB3 for 24 h) shows a NE-like staining in living human myoblasts (left panel) and in fixed (right panel) HeLa cells. DAPI staining is shown. Scale bar = 5 µm. Related to Supplementary Fig. S1.
