Fig. 6

GRIN1 and GNAO1 activate NLRP4E, which subsequently regulates SRC and CDC42 phosphorylation and actin cap assembly. A We used NLRP4E antibody to perform IP and MALDI and identified four interacting proteins, one of which was GRIN1. B Co-IP and western blotting verified that NLRP4E interacted with GRIN1. C Co-IP- and western blotting show that GANO1interacted with NLRP4E. D Immunofluorescence shows that GANO1 colocalized with NLRP4E at the oocyte membrane. NLRP4E is shown in green, GANO1 is in red, and DNA is in blue. E–H Western blots showing that both GRIN1 (E and F) and GANO1 (G and H) knockdown significantly reduced p-NLRP4E. I, J IP-blot showing that both GANO1 (I) and NLRP4E (J) interacted with SRC. K Immunofluorescence showing that NLRP4E colocalized with SRC at the oocyte membrane. NLRP4E is shown in green, SRC is in red, and DNA is in blue. L–O Western blot showing that both GANO1 (L and M) and NLRP4E (N and O) knockdown significantly reduced p-SRC. P Co-IP blot showing that SRC interacted with CDC42. Q–T Western blots showing that both NLRP4E (Q and R) and SRC (S and T) knockdown significantly increased p-CDC42. Scale bar, 20 μm.  * indicates p  < 0.05; ** indicates p  < 0.01; *** indicates p  < 0.001; **** indicates p  < 0.0001