Fig 1: Disruption of TDP-43 impacts clock genes regulation(A) Luciferase activity driven by E-box in HEK293 cells following transfection with varying amounts of TDP-43 or CRY1 expression vectors in a 96-well plate. n = 3 represents technical replicates.(B) Transient transfection luciferase assay demonstrating differential repression activity in shTDP-43 and shcon cells. n = 3 represents technical replicates.(C) mRNA levels of indicated clock genes in shTDP-43, shcon, and U2OS cells harvested every 4 h after synchronization (medium change). n = 3 samples per time point. The JTK_Cycle analysis was conducted to evaluate the rhythmicity of gene expression. JTK_p < 0.05 indicates significantly rhythmic.(D) Expression profiles of CLOCK, PER2, CRY1, and CRY2 in shTDP-43, shcon, and U2OS cells harvested every 4 h after synchronization (medium change). Bottom: JTK_Cycle analysis was conducted to evaluate the rhythmicity of gene expression. JTK_p < 0.05 indicates significantly rhythmic. One-way ANOVA with multiple comparison test for (A) and (B), two-way ANOVA with multiple comparison test for (C) and (D). Data are presented as mean ± SEM, ns indicates no significance; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.