Fig. 3: Ptbp1 is transcriptionally activated by FOXN1.

a Expression of Ptbp1 in terminal oocytes of gregarious (G) and solitarious (S) locusts (n = 5 biologically independent locusts). b-d A diagram (b), PCR results (c), and promoter activity (d) of the progressively truncated upstream sequences of ATG start codon of Ptbp1 (n = 6 biologically independent cell samples). e A diagram for binding of the predicted transcription factors (TFs) on Ptbp1 promoter. f PCR of the TFs candidates from various tissues. g-h The effects of three TFs knockdown on the expression of Ptbp1 (g) and miR-276/pre-miR-276 ratio (h) (n = 5 biologically independent locusts). i-j Expression of mRNA (i; n = 5 biologically independent locusts) and protein (j; n = 4 and five biologically independent locusts respectively for G and S) of Foxn1. k-m Validation of the binding of FOXN1 with the Ptbp1 promoter by EMSA from the nuclear extracts of terminal oocytes. Red and blue letters indicate the binding sites of wild-type (WT) probe and the mutations in mutated (Mut) probe, respectively (k). The bands of free probe and shift band are indicated by arrows (l and m). n The effects of overexpression of locust FOXN1 (top) on the Ptbp1 promoter determined by dual-luciferase assay (bottom) (n = 5 biologically independent cell samples). Rp49 (f) and GAPDH (j and n) were used as internal control for genes and proteins, respectively. Statistically significant differences between groups in (d) are shown by different letters using one-way ANOVA followed by Tukey’s post hoc tests with two-stage step-up method of Benjamini, Krieger and Yekutieli adjustment for multigroup comparisons. Mann–Whitney U (two-tailed) test was applied for two-group comparisons in (a), and Student’s t test (two-tailed) was applied for other two-group comparisons. NC cells transfected with empty vector. The data are shown as the mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001. The original scans for Fig. 3c, 3f, 3j, and 3n are provided in Supplementary Figs. 15a, 15c, 17b and 17c, respectively. Source data and details of the statistical results are provided as a Source Data file.