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. 2021 Mar 29;9:650250. doi: 10.3389/fcell.2021.650250

FIGURE 3.

FIGURE 3

Inhibition of JAK-STAT signaling pathway inhibits cellular senescence. (A,B) The transfection efficiency of JAK2-siRNA was explored by qRT-PCR and Western blotting in aged TSPCs. (C,D) The expression of STAT3 mRNA and protein was measured by qRT-PCR and Western blotting in aged TSPCs after treatment with STAT3-siRNA. (E,F) β-gal staining for the senescent cells in aged TSPCs upon JAK2-siRNA or STAT3-siRNA treatment. Scale bars: 100 μm. (G,H) The expression of p16INK4A mRNA and protein was measured by qRT-PCR and Western blotting. (I) The cell cycle distribution of TSPCs was measured by flow cytometry after JAK2-siRNA/STAT3-siRNA treatment. (J,K) β-gal staining for the senescent cells in aged TSPCs after treatment with AG490 treatment (100 μM, 24 h). Scale bars: 100 μm. (L,M) The p16INK4A level was investigated by qRT-PCR and Western blotting in young, aged, and aged AG490-treated TSPCs. (N) Cell cycle analysis of aged TSPCs after AG490 treatment for 48 h. *P < 0.05 compared with the young or aged group.