Figure 1.

Distinct point mutations in the LRBA gene of two LRBA-deficient patients cause mRNA decay and loss of the protein. (A) Schematic of LRBA protein structure with annotated domains. The genetic mutations carried by the two LRBA-deficient patients investigated in this study are shown. Dermal fibroblasts obtained from these patients were used in this study. (B) LRBA mRNA levels in the two patient and two healthy donor (HD) fibroblast cell lines were determined by qRT-PCR. Mean and standard deviation are shown from n = 4 biological replicates; one-way ANOVA using Dunnett’s multiple comparison, ****P < 0.0001. (C) Immunoblot analysis of LRBA presence in fibroblasts of two HDs and two patient donors using polyclonal LRBA antibody and actin as a loading control. (D) Colocalization of LRBA and TGN46 in fibroblasts of HDs. LRBA is absent in patient-derived fibroblasts. Cells were fixed, immunostained with TGN46 and LRBA antibodies, and imaged using a confocal microscope. Squares show magnification of the perinuclear area (1) and the periphery (2). The labeling of the single channels represents the color of the channel on the merged image. H1: HD 1, H2: HD 2, P1: patient 1, P2: patient 2. Source data are available for this figure: SourceData F1.