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. 2022 Apr 28;80:104019. doi: 10.1016/j.ebiom.2022.104019

Figure 4.

Fig 4

m6A methylation regulates the expression of regulators of STAT3 activation. a. There was no distinct effect on the STAT3 m6A modification after ALKBH5 overexpression. b. SOCS3 m6A methylation decreased after ALKBH5 overexpression. c. Mutations at the four putative m6A sites in SOCS3 (A to T) were generated. d. ALKBH5 decreased the m6A modifications of SOCS3-WTand SOCS3-MUTs (which contains only one potential m6A site) in osteosarcoma cells. And the decrease degree of m6A modification was suppressed in SOCS3-MUTs compared to SOCS3-WT.While the m6A modification of SOCS3-MUT1–4 (which contains all four potential m6A sites mutation) was not decreased in osteosarcoma cells with overexpression of ALKBH5. e. SOCS3 expression was elevated when ALKBH5 was overexpressed both in U2OS and KHOS cells. f. SOCS3 m6A modification level of osteosarcoma cells was decreased when treated with DAA. g. SOCS3 expression was augmented strongly when treated with DAA. h. The decrease of m6A modification caused by DAA was not detected in SOCS3-MUT1–4 cells. i. SOCS3 expression negatively correlated with m6A methylation in osteosarcoma tissues. The data represent the mean± S.D. of three independent experiments. ANOVA or t-tests was used for statistical analysis. *P<0.05 or **P<0.01 indicates a significant difference between the indicated groups.