Figure 3.

Rab 5 and SNX-27 form a ternary complex with IP6K2–4.1N. A, Workflow for screening proteins that bind to the IP6K2–4.1N complex. B, Immunoprecipitates (IPs) of Rab 5 proteins blotted with IP6K2 and 4.1N antibodies, respectively, revealed the binding of Rab 5 to both IP6K2 and 4.1N in wild-type mouse brain lysates. C, Immunoblot (IB) showed that SNX-27 binds to 4.1N and not to IP6K2. D, Western blot of wild-type and IP6K2-KO mice brain lysates showed reduced the protein expression of Rab5 and SNX-27 in IP6K2-KO mouse brain lysates. E, Quantitation of the relative expression of Rab 5 and SNX-27 depicted in D through normalization against the relative expression of the relevant input. F, Western blots depict reduced expression of some prominent Immediate Early Gene (IEG)-encoded proteins like Arc, Fos B, NGF, and Synaptophysin (Syn) in the IP6K2-KO mice brain lysates. G, Quantitation of the depicted. Western blots in F through normalization against the input is shown in the histogram. Data were statistically analyzed by paired Student's t test. Significant differences (*p < 0.05) were observed in comparison with WT controls. Data are presented as the mean ± SD and are representative of three independent experiments done under identical conditions.