Fig. 8

Fig. 8 Interactions between NH₂-terminus containing the RBD and the WDR are affected by patient variants.

a Immunoprecipitation of endogenous WDR44 from controls and p.S764F patient fibroblast in the presence of 10% serum (+serum) or starved (−serum) for 24 h. Immunoblots were probed with WDR44, pAkt-substrate (pAKT WDR44), PanAKT, and β-actin antibodies. Blots are represented from two independent experiments. b Immunoblot analysis of Akt phosphorylation of Myc-WDR44 wild-type or G782C or S764F, and GFP-RAB11 co-immunoprecipitated under ciliating conditions (3 h and 24 h serum starvation) from 293T cells expressing indicated plasmids for 48 h. GAPDH was used to evaluate starting lysate protein levels. Blots are represented from two independent experiments. c The schematic represents (top) WDR44 domains, HA-NH₂-terminal domain containing RBD (1-408AA), and GST-COOH-terminal domain containing WDR (477-913 AA). Immunoblotting analysis (bottom left) showing co-immunoprecipitation between HA-NH₂-terminal domain and GST-COOH-terminal domain of wild-type or variants from 293T cells expressing indicated plasmids for 48 h. Plot (right) shows the relative co-immunoprecipitated levels of GST-COOH-terminal WDR44 compared to HA-NH2-terminal WDR44 proteins normalized to pre-IP levels of GST-COOH-terminal. P = 0.0224 (N840S), 0.0035 (L668S), 0.0021 (S764F), 0.0031 (G782C), 0.0001 (D648G), 0.0004 (D669N), 0.0008 (H839R). Mean ± s.e.m. from three independent experiments. Unpaired two-tailed t-test; *P < 0.05, **P < 0.01, ***P < 0.001. d Summary of WDR44 variant results. Source data are provided as a Source Data file.