Fig. 5: PTBP1-mediated nuclear export is dependent on the “CU motif”. | Nature Communications

Fig. 5: PTBP1-mediated nuclear export is dependent on the “CU motif”.

From: Parental experiences orchestrate locust egg hatching synchrony by regulating nuclear export of precursor miRNA

Fig. 5

a Cytoplasmic/nuclear ratios of the eight selected pre-miRNAs in terminal oocytes after knockdown of Ptbp1 (n = 5 biologically independent locusts). b Sequences of wild type (WT) and mutant pre-miR-276 (Mut). Red boxes indicate the “CU motif” sequences. The red and blue letters indicate the mutant bases within the “CU motif” and “non-CU motif”, respectively. c Secondary structures of WT and Muts. d Expression of pre-miRNAs produced from the mutated plasmids (n = 5 biologically independent cell samples). e-f Effects of PTBP1 on cytoplasmic/nuclear pre-miRNA ratios in HEK 293T cells. The ratio was normalized to that of the LacZ transfection group (n = 5 biologically independent cell samples). The red line indicates that the cells transfected with PTBP1 or LacZ had the same cytoplasmic/nuclear pre-miRNA ratio. g–h miRNA/pre-miRNA ratios in cells transfected with the wild-type or mutated plasmids. The ratio was normalized to that of the LacZ transfection group (n = 5 biologically independent cell samples). The red line indicates that the cells transfected with PTBP1 or LacZ had the same miRNA/pre-miRNA ratio. RIP assay using antibodies against PTBP1 (i–j) or XPO5 (k–l) in HEK 293T cells after transfection with WT and mutated vectors (i: n = 5 biologically independent cell samples for Mut1 and Mut3 group, n = 4 biologically independent cell samples for Mut2 group; j and l: n = 5 biologically independent cell samples; k n = 4 biologically independent cell samples). The red line indicates that the cells transfected with PTBP1 or LacZ had the same abundances of pre-miRNAs in the immunoprecipitates of XPO5. Mann–Whitney U test (two-tailed) was applied for two-group comparisons of pre-miR-13a, pre-miR-100, pre-miR-92 and pre-miR-14 in (a), mut 4 in (d) and mut 6 in (h). Student’s t test (two-tailed) was applied for other two-group comparisons. The data are shown as the mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, n.s. no significant difference. Source data and details of the statistical results are provided as a Source Data file.

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