Figure 3
From: Endogenous TDP-43, but not FUS, contributes to stress granule assembly via G3BP
Blocked SG assembly in TDP-43 depleted cells is rescued by G3BP-GFP. (A - F) SG formation and resolution were assessed in HeLa cells transfected with control or TDP- 43 siRNA for 48 hr, transfected with GFP or G3BP-GFP for 24 hr and subsequently treated with SA. Cover slips were collected before SA treatment (A – C) and at 60 and 90 min after stress (D - F). (A) Western blot confirming expression of transfected plasmids. Actin serves as a loading control. (B) Representative micrographs of quantified cells with spontaneous SG formation. Asterisks and arrows indicate cells expressing G3BP-GFP with or without spontaneously forming SG, respectively. Scale bar, 20 μm. (C) Quantification of cells with spontaneous SG formation (SG formation in the absence of stress). Cells were scored as SG positive when they had at least two TIA-1 foci of a minimal size of 0.75 μm2. (D) Representative images quantified for SG number and size by semi-automatic analysis using ImageJ. Scale bar, 10 μm. (E & F) Number and size of SG were quantified at 60 and 90 min after SA treatment by semi-automatic analysis using ImageJ. The means of 3 independent experiments ± SEM are plotted. * p < 0.05.