Fig. 6

Pnt directly binds to Jhamt promoter region and induces Jhamt expression. A Western blotting confirmed the successful overexpression of Pnt in Drosophila KC cells. B–C Relative luciferase activity of Jhamt (− 2000 nt~− 1 nt) (B) and Cyp15A1 (− 3000 nt~− 1 nt) (C) promoter transfection with PIEX4-Pnt-Flag. ***P < 0.001, ns (not significant, P > 0.05), n = 3 or 4. D–D” Relative luciferase activity of different regions of Jhamt promoter. The orange line represents the Jhamt promoter fragment that cloned into the pGL3 vector, and the black line represents the Jhamt promoter excised fragment. E Relative luciferase activity of Jhamt promoter mutants. ***P < 0.001, n = 3 or 4. F EMSA analysis of the binding of nuclear proteins Pnt extracted from KC cell using a region between − 941 and − 886 nt of Jhamt promoter and the mutated probes or flag antibody. The mutant nucleotides are marked in red. G Schematic representation of the Egf ligands Vn and Spi activate transcription factor Pnt via Ras/Raf/ERK signaling to promote JH biosynthesis in the CA