Fig. 4: Silencing of dystroglycan results in altered nucleoli structure and reduced levels of B23 and UBF. | Cell Death & Disease

Fig. 4: Silencing of dystroglycan results in altered nucleoli structure and reduced levels of B23 and UBF.

From: The intracellular domain of β-dystroglycan mediates the nucleolar stress response by suppressing UBF transcriptional activity

Fig. 4

a Messenger RNA expression of dystroglycan and GAPDH (endogenous control) was assessed by qRT-PCR in C2C12 cells stably expressing DG shRNA or control shRNA, as described in Methods. Relative mRNA levels obtained in control cells were set at 1. Results are the mean ± standard deviation of three independent experiments, with P value indicating significance difference (unpaired t-test). b Lysates from C2C12 cells stably transfected with DG shRNA or control shRNA were analyzed by western blotting using primary antibodies against DG, B23, fibrillarin, UBF, and calnexin (loading control). Blots were stripped and reprobed successively with each protein antibody. Relative protein levels from control shRNA cells were set at 1 for comparison. Results are the mean ± SEM of three independent experiments (right panel), with p value denoting significance differences (unpaired t-test). c, d Nucleolar morphology was analyzed in cells stably expressing DG shRNA or control shRNA, using specific antibodies against UBF and B23 respectively. Cells were counterstained with DAPI to visualize nuclei prior to CLSM analysis. The nucleolar area (μm2) was determined in each cell condition as described in Methods (graphs at right). Data are the mean ± SEM from three independent experiments (n = 600 nucleoli), with p values indicating significant differences (unpaired t-test)

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