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

Fig. 8. Inhibiting the TRAF4/ Bcl-xL axis overcomes radioresistance in colorectal cancer cells.

Fig. 8

A–C Parental and radioresistant HT29 cells expressing sg-Ctrl or sg-TRAF4 were treated with or without IR (4 Gy). Cell viability and colony formation were determined by MTS (A), plate colony formation (B), and soft agar (C) assays. P, parental cells; R, resistant cells. ***p < 0.001. D–G HT29R cells were treated with A-1155463 inhibitor (2 µM), IR (4 Gy), or a combination of both for 72 h. MTS assay was used to determine the cell viability (D), plate colony formation assay to analyze the cell proliferation (E), and soft agar assay to assess the anchorage-independent cell proliferation (F). Immunoblotting to detect the expression of cleaved-caspase 3 and cleaved-PARP (G). ***p < 0.001. H. Average tumor volume of HT29R-sgCtrl and -sgTRAF4 xenograft tumors treated with or without IR (2 Gy/twice per week); n = 5 mice per group. I-K. Xenograft tumors from HT29R cells were treated with the vehicle control, A-1155463 (5 mg/kg/every 2 days), IR (2 Gy/twice per week), or a combination of both; n = 5 mice per group. Average tumor volume (I) and tumor weight (J) were recorded, and tumor sections were subjected to IHC staining of Ki67 and cleaved-caspase 3 (K). For K, left, representative image; right, qualification. ***p < 0.001. Scale bar, 25 μm. TRAF4 TNF receptor-associated factor 4, IR ionizing radiation, Ctrl control, P parental cells, R resistant cells.