Figure 4.

Loss of Arhgef2 inhibited the translation of m⁶A-modified Npdc1 mRNA and nuclear export of m⁶A-modified Cend1 mRNA

(A–D) mRNA expression of Npdc1 and Cend1 showed no significant change determined by qRT-PCR. Protein levels of Npdc1 and Cend1 apparently decreased in Arhgef2−/− mice compared with wild-type ones. n.s.: no significant difference. n = 4–6 per genotype, ∗p < 0.05, ∗∗p < 0.01, unpaired t test.

(E) Npdc1 mRNA expression in polysome fractions significantly reduced in Arhgef2−/− mice compared with wild-type ones, but Cend1 mRNA showed no change. Poor expression of lncRNA (Malat1 and Neat1) in polysome fraction indicated the successful extraction of polysome fraction from other fractions. n.s.: no significant difference. ∗p < 0.05, ∗∗∗p < 0.001, unpaired t test. Total: total RNA from brain tissue. Polysome: polysome fraction RNA.

(F) Compared with wild-type mice, the nuclear Cend1 level, but not Npdc1, was significantly elevated in Arhgef2−/− mice. U1 was used as control gene in nuclei. Only low β-actin expression (~5-fold cytoplasm increase over nuclear) represented the successful separation of nuclear fractions. ∗∗p < 0.01, ∗∗∗p < 0.001, unpaired t test. n.s.: no significant difference. Cyto: cytoplasm. Nuc: Nuclei. Data are represented as mean ± S.D.