Fig. 3: C3ORF52 association with lipase H.
From: Loss-of-function variants in C3ORF52 result in localized autosomal recessive hypotrichosis
(a) Normal skin scalp biopsies were stained with an anti-C3ORF52 antibody (green) and with an anti-lipase H antibody (red). Both proteins were observed to partially colocalize along the inner root sheath area (merge) (scale bar, 10 μm; blue staining, DAPI); negative controls are shown in Fig. S4. (b) Coimmunoprecipitation (Co-IP) assay using proteins extracted from HeLa cells cotransfected with LIPH and with V5-tagged C3ORF52 (wild-type [WT] or mutated [MUT, carrying the p.Tyr164Ter variant]). Protein extracts (input) were immunoprecipitated with a mouse monoclonal anti-V5 epitope tag antibody bound to protein G SureBeads and resolved by western blot. Protein–protein interactions were immunodetected using an anti-LIPH antibody. Protein blot shows a signal at about 51 kD, corresponding to the size anticipated for the lipase H protein (indicated by a red arrow). Cell lysates from HeLa cells transfected with either a pcDNA3.1 empty vector (EV) or with the wild-type or the mutated C-terminus V5-tagged C3ORF52 alone served as controls. The experiment was repeated twice with similar results. The original coimmunoprecipitation images are shown in Fig. S5.
