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. 2019 Nov 6;19(1):360–370. doi: 10.1021/acs.jproteome.9b00575

Figure 1.

Figure 1

TDP-43 knockdown leads to chromatin instability. (A) By western analysis, TDP-43 protein levels were found to be lower in HEK293 cells after targeted by two siRNAs: siTDP-1 and siTDP-2. Knockdown of TDP-43 did not affect the FUS expression, nor did FUS knockdown with siFUS affect TDP-43 protein levels. Knockdown results were compared with those after transfection of an siRNA with a scrambled sequence, SCR. (B) Comet tail assays were performed for SCR, siTDP-1, siTDP-2, and siFUS-treated HEK293 cells after treatment with etoposide or DMSO as a vehicle control. Comet tails comprising fragmented DNA were observed for siTDP-1 and siTDP-2 knockdowns after DMSO or 5 μM etoposide treatment for 1 h. Comet tails were also observed after etoposide treatment of cells with FUS knocked down with siFUS. (C) For each treatment, the percentage of cells with comet tails was counted from 40 to 140 cells and three biological replicates. Error bars indicate the standard error about the mean of replicates. * indicates p < 0.05 and ** indicates p < 0.005 with Student’s t-test assuming equal variances and compared to the SCR-treated samples.