Skip to main content
. 2021 Mar 22;3(1):zcab010. doi: 10.1093/narcan/zcab010

Figure 1.

Figure 1.

TRDMT1G155V is highly ubiquitinated in cells. (A) The survival rate of MCF-7, U2OS and SKOV3 cells with siCtrl or siTRDMT1 and indicated dose of cisplatin (n = 3, mean ± SD). (B) Schematic diagram of TRDMT1 and TRDMT1G155V mutant. (C) WB ofTRDMT1 in GFP-tagged TRDMT1WT, TRDMT1C79A orTRDMT1G155V stably expressed TRDMT1 KO 293 cells. (D) The ubiquitination level of TRDMT1 in GFP-vector TRDMT1WTor GFP-TRDMT1G155Vtransfected TRDMT1 KO 293 cells. Indicated cell lines were treated with or without 20 μM MG132 for 6 h before protein extraction. (E) WB of GFP-TRDMT1 in TRDMT1WTor GFP-TRDMT1G155V transfected TRDMT1 KO U2OS cells treated with 100 μg/ml CHX with or without 20 μM MG132 for the indicated time. (F) Dot blot analysis of m5C in mRNA of TRDMT1 KO 293 cells with stable expression of GFP-tagged TRDMT1WT, TRDMT1C79A or TRDMT1G155V. (G–H) γ-H2AX staining in GFP-tagged TRDMT1WT or TRDMT1G155Vtransfected TRDMT1 KO U2OS-TRE cells. Cells were irradiated with 6 Gy IR and stained with γ-H2AX at the indicated time point, numbers of γ-H2AX foci in each cell (G) and the frequency of 100 cells with γ-H2AX foci (H) were counted in each experiment (n = 3, mean ± SD). (I) The survival rate of the cells after IR or Cisplatin in GFP-tagged TRDMT1WT or TRDMT1G155VtransfectedTRDMT1 KO U2OS-TRE cells (n = 3, mean ± SD). Statistical analysis was done with the Student's t-test, *: P< 0.05; **: P< 0.01; ***: P< 0.001; ****: P< 0.0001 for all the figures in the paper.