Fig. 1: Functional characterization of GPATCH11 variants identified as causative for syndromic inherited retinal disease and clinical features of affected individuals.
From: GPATCH11 variants cause mis-splicing and early-onset retinal dystrophy with neurological impairment
a Pedigrees of six analyzed families, including segregation analysis of GPATCH11 variants: M1: c.328+1G>T (p.Gly97_Thr110del); M2: c.454C>T (p.Arg152*); M3: c.449+1G>C (p.Arg150ArgFs*1); M4: c.393C>G (p.Tyr131*); +: wild-type allele; P: patient. P1 and P2 are affected individuals whose fibroblasts were analyzed. b Clinical images of affected individuals P1, P2, P5, P8, P9, P11, P12. c Color fundus photographs and fundus autofluorescence images of P10, RE: right eye; LE: left eye. d GPATCH11 diagram indicating the positions of c.328+1G>T, c.454C>T, c.449+1G>C, and c.393C>G variants relative to NM_174931.4 transcript. e Representative chromatograms of gDNA sequences from affected individuals show variants: c.328+1G>T (M1) homozygously (as in P1-P4, P8, P9, P12), c.454C>T (M2) in exon 6 heterozygously (as detected in compound heterozygosity with M1 in P5-P7), c.449+1G>C (M3) homozygously (P10), and c.393C>G in exon 5 in homozygosity (P11). f Chromatograms of cDNA sequences reveal a shorter transcript (r.287-328del) in P1 and P2 due to M1, absence of transcripts (r.0) in P5 (M2), and two distinct transcripts (r.449+128 and r.449+48) in P10 caused by M3. Electrophoresis of PCR products from cDNA from P1 and control individual (C1) shows exclusion of exon 4 (42 base pairs, bp), leading to creation of a shorter aberrant transcript. Electrophoresis of RT-PCR products from leukocytes of control (C3) and P10, and schematic representation showing two aberrant transcripts present for P10 and absent in C3, corresponding to the retention of full and partial intron 5. Blots are representative of three independent experiments. g Western blot analysis of protein extracts using anti-GPATCH11 antibody targeting residues 111-192 showing detection of shorter protein isoform (GPATCH11-Δex4 (36 KDa)) in patients’ fibroblasts (P1, P2) as compared to the controls (GPATCH11-WT (37 KDa)) in C1 and C2. The abundance of GPATCH11 products in fibroblasts from affected individuals is comparable to control samples. The statistical significance was estimated using one-way ANOVA with a post-hoc Tukey test. Bars represent means ± SEM from three experimental replicates (five technical replicates). n.s. not significant. Source data are provided as a Source Data file.
