Fig 1: TRMT112 and MTases mutually stabilise each other’s expression in cells. (A) TRMT112 stabilises the expression of partner MTases in cells. The EGFP-fused MTases were expressed with and without TRMT112-E2Tag in U2OS cells, and their expression was analysed by Western blot using antibodies against EGFP (to detect MTases), E2Tag (recombinant TRMT112), TRMT112 (both endogenous, marked with asterisk, and recombinant TRMT112, upper band) and β-tubulin (loading control). (B) Flow cytometry analysis of the expression level of EGFP-fused proteins after co-expression with TRMT112. Mean fluorescent intensity (MFI) ratios are calculated as relative values of methyltransferase expression with and without TRMT112. EGFP alone was set to 1. Average of three biological replicates are shown. The difference in the sample value compared to control sample value is statistically significant with p-value * < 0.05, ** < 0.01 and *** < 0.001. (C) TRMT112 protein expression is stabilized by co-expression of its partner MTases. Cells were analysed 48 h after transfection with Western blotting using antibodies against EGFP, TRMT112 and tubulin. Mock-cells without added expression vectors. (D) Increasing TRMT112 protein level in cells does not affect the endogenous level of its partner MTases. U2OS cells were transfected with increasing amounts of TRMT112-E2Tag plasmid construct and cells were analysed 48 h later with Western blotting using antibodies against TRMT112, E2Tag, WBSCR22, N6AMT1, METTL5, ALKBH8 and tubulin (50 kDa, loading control).