Raw Western blot images for "Patient-derived organoids reveal a spliceopathy-metabolism axis underlying PRPF31-mutant retinal degeneration and therapeutic targets"

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 S4C): Relative PRPF31 protein levels in in WT versus PRPF31 c.238+1G>T engineered (WTmut) hiPSCs, normalized to VINCULIN. Raw Data Figure 4 (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 5 (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 6 (Corresponding to Manuscript Figures 6A): Relative HIF1A expression in RP11#1 versus IC retinal organoids, normalized to VINCULIN. Raw Data Figure 7 (Corresponding to Manuscript Figures 6G): Relative TSGA10 expression in RP11#1 versus IC retinal organoids, normalized to VINCULIN. Raw Data Figure 8 (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 9 (Corresponding to Manuscript Figure S7J): GAPDH, PKM2, and LDHA protein levels in ARPE19 cells expressing shPRPF31 versus control, normalized to VINCULIN. Raw Data Figure 10 (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 11 (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.本数据集包含支撑论文《患者来源类器官揭示PRPF31突变所致视网膜变性的剪接异常-代谢轴及治疗靶点》的原始蛋白质印迹（Western Blot）图像。 内容概述： 原始数据图1（对应论文图1C）：野生型（WT）与患者来源人类诱导多能干细胞（hiPSCs，RP11#1、RP11#2、RP11#3）中PRPF31蛋白相对水平，以纽蛋白（VINCULIN）为内参进行归一化处理。 原始数据图2（对应论文图1F）：野生型（WT）、RP11#1及CRISPR校正的同基因对照（IC）中PRPF31表达相对水平，以纽蛋白（VINCULIN）为内参进行归一化处理。 原始数据图3（对应论文补充图S4C）：野生型（WT）与经PRPF31 c.238+1G>T位点工程改造的突变型（WTmut）人类诱导多能干细胞中PRPF31蛋白相对水平，以纽蛋白（VINCULIN）为内参进行归一化处理。 原始数据图4（对应论文图4B）：RP11#1视网膜类器官经2-脱氧葡萄糖（2-DG）处理后，GAPDH与PKM2蛋白相对水平，以纽蛋白（VINCULIN）为内参进行归一化处理。 原始数据图5（对应论文图4I）：野生型（WT）视网膜类器官经果糖-1,6-二磷酸（F1,6BP）补充培养后，GAPDH与PKM2蛋白相对水平，以纽蛋白（VINCULIN）为内参进行归一化处理。 原始数据图6（对应论文图6A）：RP11#1与同基因对照（IC）视网膜类器官中缺氧诱导因子1α（HIF1A）表达相对水平，以纽蛋白（VINCULIN）为内参进行归一化处理。 原始数据图7（对应论文图6G）：RP11#1与同基因对照（IC）视网膜类器官中睾丸特异性基因10蛋白（TSGA10）表达相对水平，以纽蛋白（VINCULIN）为内参进行归一化处理。 原始数据图8（对应论文补充图S6H）：来自同基因对照（IC）与RP11#1的诱导性视网膜色素上皮细胞（iRPE）中PRPF31、GAPDH、PKM2及LDHA蛋白相对水平，以纽蛋白（VINCULIN）为内参进行归一化处理。 原始数据图9（对应论文补充图S7J）：表达靶向PRPF31短发夹RNA（shPRPF31）的人视网膜色素上皮细胞系ARPE19细胞与对照细胞中GAPDH、PKM2及LDHA蛋白水平，以纽蛋白（VINCULIN）为内参进行归一化处理。 原始数据图10（对应论文图6I）：人视网膜色素上皮细胞系ARPE19细胞中的遗传互作研究： PRPF31单基因敲除（KO） PRPF31敲除联合TSGA10过表达（OE） PRPF31敲除+TSGA10过表达+TSGA10敲除 检测标志物：TSGA10、PKM2、LDHA、GAPDH及PRPF31，以纽蛋白（VINCULIN）为内参进行归一化处理。 原始数据图11（对应论文图7G）：腺嘌呤碱基编辑器（ABE）介导的碱基编辑对RP11#3视网膜类器官中PRPF31蛋白表达的挽救效果，以纽蛋白（VINCULIN）为内参进行归一化处理。 注：每张图像左侧标注了分子量标志物。泳道标识与论文主图中的样品标签一致。用于内参对照的抗体包括抗纽蛋白（VINCULIN）抗体（Abcam，货号ab129002，稀释比例1:20000；Abmart，货号T40106，稀释比例1:5000）。关于细胞培养、转染及药物处理的完整实验流程详见相关论文的方法部分。