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. 2021 Apr 21;9:623394. doi: 10.3389/fcell.2021.623394

FIGURE 4.

FIGURE 4

Nsun5 regulated Tpm1 as a m5C methyltransferase. (A) The mRNAs isolated from E14.5 Nsun5 hearts were used in dot blot analyses with m5C antibody. n = 3/group. (B) GO-BP analysis for the DMSs genes in E14.5 Nsun5+/+ and Nsun5– /– hearts. (C) GO-BP analysis for the differentially expressed proteins in E14.5 Nsun5+/+ and Nsun5– /– hearts. (D) The overlap of identified DMSs genes and differentially expressed proteins by BS-seq and LC-MS/MS in Nsun5– /– E14.5 hearts when compared with Nsun5+/+ controls. (E) DMSs in Tpm1. Fold change of each DMS was indicated by the node size. P-value of each DMS was indicated by the bar color. The black horizontal line represents P = 0.05. (F) The protein expression of Tpm1 confirmed by western blot in E14.5 Nsun5 hearts. n = 3/group. (G,H) Quantitative results showed reduced Tpm1 intensity in Nsun5 deletion OFT aorta area and ventricular septum by immunofluorescence staining. The data was presented as the ratio of Tpm1+ area intensity/total area intensity. Cardiomyocytes were labeled by TnT. n = 5/group. Scale bar = 50 μm. All the data were the mean ± SEMs of three dependent experiments. One-way ANOVA test and hypergeometric test were used for statistical calculation. *P < 0.05, **P < 0.01.