Fig. 5

KIAA1429 reduced the stability of CHST11 mRNA in an m6A-YTHDF2-dependent manner. A Visualization of m6A peak in CHST11 transcripts based on MeRIP-seq, and reduction of m6A levels of CHST11 by knockdown of KIAA1429. B Construction of a luciferase reporter containing wild-type CHST11 3′ UTR and CHST11 3′ UTR with a mutation at the m6A consensus sequence. C, D An analysis of relative luciferase activity was conducted in DLBCL cells with or without KIAA1429 knockdown or KIAA1429 overexpression. Triplicate data are shown as mean ± SD. **p < 0.01; ***p < 0.001. E, F RT-qPCR was used to detect the expression of CHST11 after treatment with actinomycin D (5 μg/ml) at the indicated timepoints in KIAA1429 knockdown, KIAA1429-overexpressing, and corresponding control DLBCL cells. Result are presented as mean ± SD from three independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. G, H Knockdown efficiency of CHST11 in DLBCL cells was verified at mRNA and protein levels. As a result of three independent experiments, mean ± SD is presented. *p < 0.05; ***p < 0.001. I RT-qPCR was utilized to determine the alterations in expression of m6A-binding proteins upon CHST11 knockdown. The results are shown as mean ± SD from three independent experiments. **p < 0.01; ****p < 0.0001. J The relative mRNA levels of CHST11 were detected in DLBCL cells with YTHDF2 overexpression or knockdown. Data from three independent experiments are presented as mean ± SD. *p < 0.05; **p < 0.01; ***p < 0.001. K Validation of direct binding between YTHDF2 and CHST11 mRNA was performed by RIP-qPCR. Data are expressed as mean ± SD from three independent experiments. **p < 0.01; ***p < 0.001; ****p < 0.0001