TDP-43 regulates the expression of USP13. (A) USP13 was found from two sets of RNA-seq data (Roczniak-Ferguson and Ferguson, 2019). (B) HEK-293T cells were transfected with PX458/TDP-43KO (TKD) and HA-TDP-43 24 h later. The cells were collected 48 h later and RNA was extracted and performed to RT-qPCR. ns, no significance. (C) HEK-293T cells were transfected with pLL3.7, shTDP-431, and shTDP-432. The cells were collected at different CT and analyzed using western blotting and quantitated by comparing by two-way RM ANOVA. (D) M17 cells were transfected with PX458/TKD, and RNA was extracted 48 h later. The splicing products of USP13 were analyzed using RT-PCR and the ratio of P3/P1 was calculated. (E) HEK-293T cells were transfected with shUSP13 for 48 h, and the cell lysates were analyzed using western blotting and quantitated. (F) HEK-293T cells were transfected with 0, 0.5, 1, 1.5, and 2 μg His-USP13 for 48 h, and the cell lysates were analyzed using western blotting, with the relative expression of BMAL1 quantitated in each treatment. (G) HEK-293T cells were transfected with shTDP-43 for 24 h and then transfected with His-USP13 and treated with MG132 (10 μM) for 12 h. The cells were immunoprecipitated with anti-BMAL1 and analyzed using western blotting. Data are presented as mean ± S.D. (n = 3). The unpaired t test was used in comparisons of B and D–F. **P < 0.01; ***P < 0.001; ****P < 0.0001. ns, no significance. Source data are available for this figure: SourceData F5.