Fig. 5

FKA inhibition of the methylation activity of PRMT5. a Fluorescent report CETSA assay confirmed that 25 μM FKA treatment inhibited PRMT5 degradation when heated, thus enhancing PRMT5 stability in T24. b Precipitation of H2A (left) and H4 (right) with PRMT5 and the inhibition of histone binding with PRMT5 after adding FKA to T24. c Methylation change on the third arginine of histone 2A and histone 4 when supplied with different concentrations of FKA and SAM in T24 (left) and UMUC3 (right) cells. d PRMT5 functional site mutant plasmids were induced in T24 cells, then a pull-down assay using biotin-labeled FKA was performed to determine the potential binding sites between FKA and PRMT5. e Fluorescent report CETSA assay performed for PRMT5 expression when treated with FKA in wild-type and Y304A/ F580A mutated PRMT5 in T24 cells. Data are presented as the mean ± SD of three replicates. f Bio-layer interferometry detected the combination and dissociation of FKA and Y304A mutated recombinant PRMT5. g Bio-layer interferometry detected the combination and dissociation of FKA and F580A mutated recombinant PRMT5. h Arginine methylation changed at the third arginine of histone 2A, and histone 4 in Y304/F580 mutated PRMT5 supplied with FKA and SAM in T24 cell