Fig. 4
Internal mRNA m7G promotes Sptbn2 stability and translation. (A) Integrative Genomics Viewer (IGV) tracks displaying m7G reads from NSCs, neurons and astrocytes. (B-E) Immunofluorescence staining, Western blotting and RT-qPCR of Sptbn2 expression in NSCs, neurons and astrocytes. Scale bar, 150 μm, n = 3, ***P < 0.001, **P < 0.01, *P < 0.05 compared with NSCs. (F-I) The expression of Sptbn2 protein and mRNA in NSCs transfected with the depletion or overexpression of Mettl1. n = 3, ***P < 0.001 compared with control. (J, K) Sptbn2 mRNA stability was determined by qRT-PCR in control and NSCs transfected with the depletion or overexpression of Mettl1 using the samples treated with ActD at the indicated times. n = 3, *P < 0.05 compared with control. (L) Polysome profiling of NSCs with or without Mettl1 knockdown. Data are represented as the mean ± SEM. NSCs, neural stem cells; Sh-Mettl1, shRNA Mettl1; OE-Mettl1, overexpressing Mettl1; ActD, actinomycin D; n represents number of independent experiments