Fig. 3

DCTN1 knockdown delays the disassembly of stress granules in heat-stressed cells. a Microscopic images of HEK293 cells after heat shock (HS) at 42 °C for 60 min. Upon heat stress, TDP-43 is colocalized with G3BP1, a marker of stress granules. Arrows indicate stress granules. b, c Western blot images (b) and bar graphs (c) showing relative expression levels of DCTN1 and TDP-43 in HEK293 cells that were transfected with an siRNA against DCTN1 (siDCTN1) or a non-targeting control siRNA (siControl). d Schematic representation of the HS experiments. e, f Microscopic images (e) and bar graphs (f) of stress granule formation after 30-min and 60-min HS. In (e), the panels at the bottom are magnified images of the boxed region in the top panels. In (f), the ratio of cells with cytoplasmic granules that are positive for both TDP-43 and G3BP1 were calculated. g Schematic representation of the recovery experiments after HS. h, i Microscopic images (h) and bar graphs (i) of stress granules analyzed at the recovery time of 30-min and 60-min after HS. Statistical analyses in c, f, and i were performed to assess differences between groups by the Student t-test (**p < 0.01, ****p < 0.0001; n.s., not significant). Scale bars, 10 µm (a), and 50 µm (top) and 10 µm (bottom) (e, h)