Fig. 6: Arl4A interacts with the WHB ___domain of VPS36.

a Schematic representation of VPS36 and its alanine mutants scanned from the C-terminus to a.a. 327 on VPS36 (201–386). Red letters indicate the amino acids mutated to alanine (upper). The yeast reporter strain L40 was transformed with constructs encoding VPS36 mutants fused to Gal4 AD and constructs encoding Arl4A T51N fused to LexA BD. b Purified His-tagged Arl4A T51N was incubated with the indicated GST fusion protein. Bead-bound His-Arl4A T51N was probed using an anti-His antibody. Arrows indicate the primary bands of GST-VPS36 (201–386). The amounts of Arl4A were determined by densitometric quantification and presented as the mean ± SD of four independent experiments (the p-values were assessed by ANOVA with Tukey’s test). c Conservation of amino acid ³⁵¹LAKER³⁵⁶ at the A6 region in different species. The amino acid sequence alignment of the A3 to A6 regions at the VPS36 WHB ___domain in different species as indicated was analyzed by the PRALINE multiple sequence alignment tool. d Wild-type VPS36 or the VPS36-A6 mutant interacted with Arl4A in vivo. Lysates of HeLa cells transfected with the indicated plasmids were immunoprecipitated with Myc-Trap, and the bound proteins were analyzed by Western blotting with anti-Myc, anti-Arl4A, or anti-α-tubulin antibodies. The amounts of coimmunoprecipitated Arl4A were determined by densitometric quantification and presented as the mean ± SD of three independent experiments (the p-value was assessed by a two-sided t-test). Source data are provided as a Source Data file.