Research Paper Volume 12, Issue 21 pp 22174—22198

Nucleolar stress induces a senescence-like phenotype in smooth muscle cells and promotes development of vascular degeneration

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Figure 4.

Nucleolar stress caused a senescence-like phenotype in murine vascular SMCs (MOVAS). (A) CX-5461 treatment (0.5 μM for 5 days) increased the number of β-galactosidase (β-Gal) positive cells (blue color). The numeric data were shown on top. (B) Real-time PCR results showing the expression levels of plasminogen activator inhibitor (PAI)-1 and p21Cip1 in control and CX-5461-treated (0.5 μM for 48 hr) cells (n = 5). (C) Real-time PCR results showing the effects of TIF-IA-targeting short hairpin RNA (shTIF) on the expressions of TIF-IA and unrelated Pol I components (n = 6 - 7). (D) Changed morphology of shTIF-IA-treated cells comparing to control cells. (E) Proliferation of control and shTIF-IA-treated cells assessed by CCK-8 assay (n = 3). (F) Changes in the mRNA levels of PAI-1 and p21Cip1 in shTIF-IA-treatment cells (n = 5 - 8). (G) Flow cytometry data showing G2/M blockade and S phase delay in shTIF-IA-treated cells. Data were expressed as mean ± S.D. * P < 0.05 versus control (Con), unpaired t-test. Flow cytometry assays were repeated at least 3 times.