Figure 2.

NF-κB signaling is activated during RCT development

(A) Western blotting for NF-κBp-p65 and NF-κBp65 and (B) quantification conforming the significantly increased NF-κB activation in S-TSPCs than that in H-TSPCs (n = 3 per group). (C) Immunofluorescence (green) staining and (D) quantification for NF-κBp65 revealing significantly increased nuclear NF-κBp65 localization in S-TSPCs than that in H-TSPCs. The red triangle indicates nuclear localization of NF-κBp65 (n = 3 per group). (E) Western blotting for NF-κBp-p65 and NF-κBp65 and (F) quantification conforming that IKKβ siRNA can significantly decrease NF-κB activation in IL-1β-induced senescent TSPCs (n = 3 per group). (G) Immunofluorescence (green) and (H) quantification for NF-κBp65 revealing that IKKβ siRNA can significantly decrease IL-1β-induced nuclear NF-κBp65 localization. The red triangle indicates nuclear localization of NF-κBp65 (n = 3 per group). (I) SA-β-gal staining and (J) quantification of senescent human TSPCs (n = 3 per group). (K) Immunofluorescence staining and (L) quantification of p16^INK4A (red) and p21^CIP1 (green) (n = 3 per group). NF-κBp-p65, phosphorylated NF-κB p65. NF-κBp65, unphosphorylated NF-κB p65. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.