Raw Western blot images for "Patient-derived organoids reveal a spliceopathy-metabolism axis underlying PRPF31-mutant retinal degeneration and therapeutic targets"
Description
This dataset contains the raw western blot images supporting the manuscript "Patient-derived organoids reveal a spliceopathy-metabolism axis underlying PRPF31-mutant retinal degeneration and therapeutic targets". Content Overview: Raw Data Figure 1 (Corresponding to Manuscript Figure 1C): Relative PRPF31 protein levels in WT and patient-derived hiPSCs (RP11#1, RP11#2, RP11#3) normalized to VINCULIN. Raw Data Figure 2 (Corresponding to Manuscript Figure 1F): Relative PRPF31 expression in WT, RP11#1, and the CRISPR-corrected isogenic control (IC), normalized to VINCULIN. Raw Data Figure 3 (Corresponding to Manuscript Figure 4B): Relative GAPDH and PKM2 protein levels in RP11#1 retinal organoids following 2-DG treatment, normalized to VINCULIN. Raw Data Figure 4 (Corresponding to Manuscript Figure 4I): Relative GAPDH and PKM2 protein levels in WT retinal organoids following fructose-1,6-bisphosphate (F1,6BP) supplementation, normalized to VINCULIN. Raw Data Figure 5 (Corresponding to Manuscript Figures 6A): Relative HIF1A expression in RP11#1 versus IC retinal organoids, normalized to VINCULIN. Raw Data Figure 6 (Corresponding to Manuscript Figures 6G): Relative TSGA10 expression in RP11#1 versus IC retinal organoids, normalized to VINCULIN. Raw Data Figure 7 (Corresponding to Manuscript Figure S6H): Relative protein levels of PRPF31, GAPDH, PKM2, and LDHA in induced retinal pigment epithelium (iRPE) cells derived from the IC and RP11#1, normalized to VINCULIN. Raw Data Figure 8 (Corresponding to Manuscript Figure S7J): GAPDH, PKM2, and LDHA protein levels in ARPE19 cells expressing shPRPF31 versus control, normalized to VINCULIN. Raw Data Figure 9 (Corresponding to Manuscript Figure 6I): Genetic interaction studies in ARPE19 cells: PRPF31 single knockout (KO) PRPF31 KO with TSGA10 OE PRPF31 KO + TSGA10 OE + TSGA10 KO Probed markers: TSGA10, PKM2, LDHA, GAPDH and PRPF31, normalized to VINCULIN. Raw Data Figure 9 (Corresponds to Manuscript Figure 7G): ABE-mediated base editing rescue of PRPF31 protein expression in RP11#3 Retinal organoids, normalized to VINCULIN. Note: Molecular weight markers are indicated on the left of each image. Lane identities correspond to the sample labels in the main manuscript figures.Antibodies used for loading controls included anti-VINCULIN (Abcam, Cat# ab129002, 1:20000; Abmart, Cat# T40106, 1:5000). Full experimental procedures regarding cell culture, transfection, and pharmacological treatments are detailed in the Methods section of the associated manuscript.
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Institutions
- Shanghai Jiao Tong UniversityShanghai, Shanghai