Fig. 4.

Thoc5 regulates the export of Trincr1 to cytoplasm. a Fraction of Trincr1 in cytoplasm (Cyto) and nucleus (Nuc). n = 3 independent experiments. Data were normalized by using comparable amounts of nuclear and cytoplasmic lysates from the same cell samples. U1 and Gapdh were used for the quality control of nucleus and cytoplasm fraction, respectively. b RT-qPCR analysis of knockdown efficiency of CRISPRi constructs targeting various components of transcription-export complex. The β-actin gene was used as a control. Data were normalized to ESCs treated with control gRNAs. n = 3 biological replicates. c Fraction of Trincr1 in cytoplasm and nucleus in CRISPRi ESCs. n = 3 independent experiments. Data were normalized by using comparable amounts of nuclear and cytoplasmic lysates from the same cell samples. U1 and Gapdh were used for the quality control of nucleus and cytoplasm fraction, respectively. d Western blotting analysis of phosphorylated ERK in control and Thoc5 CRISPRi ESCs in 2i + LIF. For quantification of pERK/ERK, data were normalized to GAPDH and then to control gRNA treated ESCs cultured in 2i + LIF. n = 4 independent experiments. Shown are mean ± SD. For c, P values were determined by unpaired two-way ANOVA with two-sided Dunnett’s test. For d, P values were determined by paired two-sided Student’s t-test