Fig. 4

DCTN1 knockdown promotes the formation of ubiquitin-positive TDP-43 inclusions in heat-stressed cells. a Schematic representation of the experiment. HEK293 cells that were transfected with non-targeting control siRNA or DCTN1 siRNA were heat-shocked at 42 °C for 1 h, and then incubated at 37 °C for 6 h in the presence or absence of MG-132. b Microscopic images of HEK293 cells that were treated with MG-132 for 6 h after HS. c, d Microscopic images (c) and a bar graph (d) showing cytoplasmic TDP-43 inclusions costained with the ubiquitin antibody. Arrows in b and c indicate ubiquitin-positive inclusions of TDP-43 in the cytoplasm. Statistical analysis was performed to assess differences between groups by the Student t-test (**p < 0.01). Scale bar, 20 µm (b, c)