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. 2023 Feb 10;14(2):102. doi: 10.1038/s41419-023-05637-y

Fig. 4. c-Jun is required for IR-induced Bcl-xL expression.

Fig. 4

A and B HT29 cells were transfected with sic-Jun, sip65, siSTAT5 and siEst1 for 24 h, followed by IR (4 Gy) treatment and cultured for 4 h. IB was used to examine the protein expression (A); the mRNA levels of Bcl-xL was examined by RT-qPCR (B). ***p < 0.001. C TRAF4-null HCT116 and -HT29 cells were treated with or without IR (4 Gy) and cultured for 30 min. IB was used to examine the protein expression. D TRAF4-null HT29 cells were transfected with pGL3 basic or pGL3 AP1 for 48 h and treated with or without IR (4 Gy). Reporter activity was examined 30 min later. ***p < 0.001. E and F TRAF4-null HT29 cells were transfected with Flag-TRAF4 for 48 h, followed by IR (4 Gy) treatment. Reporter activity (E) and protein expression (F) were examined 30 min later. ***p < 0.001. G. TRAF4-null HT29 cells were treated with cycloheximide for various time points, and IB was used to examine the protein expression. H TRAF4-null HT29 cells were pretreated with MG132 for 6 h, followed by IR (4 Gy) treatment for 30 min, and WCE was subjected to ubiquitination analysis. I TRAF4-null HT29 cells were overexpressed with c-Jun for 48 h, and WCE was subjected to IB analysis. TRAF4 TNF receptor-associated factor 4, IR ionizing radiation, IB immunoblotting, siRNA small interfering RNA, AP-1 activator protein-1, WCE whole-cell extract.