Fig. 2

Loss of Wdr76 increases Ras protein levels and exacerbates CRC tumorigenesis. a Kaplan-Meier analysis of overall survival in colon cancer patients, classified by WDR76 expression (WDR76 high and WDR76 low groups are defined as those with FPKM values above the 66th percentile and below the 33rd percentile, respectively). Log-rank (Mantel-Cox) test, p = 0.0441. b Linear regression curve showing LGR5 and WDR76 expression levels in log₂ in 104 CRC patients (GSE21510). c Confocal immunofluorescence of Wdr76 and Lgr5-GFP in Apc^Min/+; Lgr5-EGFP intestinal sections with linear regression curve showing Lgr5 and Wdr76 expression in tumors. Quantitative expression levels based on at least 10 tumors per 10 fields of view. *** p < 0.001. Wdr76 (red), Lgr5-GFP (green), and DAPI (blue). Boxes indicate the enlarged areas. Tumor areas are indicated by dotted lines. T: Tumor; NT: Non-tumor. Scale bars represent 20 μm. d-g Analysis of small intestinal sections of 15-week-old Wdr76^+/+; Apc^Min/+and Wdr76^−/−; Apc^Min/+ mice d H&E stained sections of small intestines from Wdr76^+/+; Apc^Min/+and Wdr76^−/−; Apc^Min/+ mice. Tumors are indicated by arrowheads. Scale bars represent 20 μm. e Quantification of the total number and size (pixel) of tumors per mouse (Wdr76^+/+, n = 6; Wdr76^−/−, n = 6). **p < 0.01. f Confocal immunofluorescence of Wdr76, Ras, PCNA, CD44, CD133, and CD166 in sections of small intestine from Wdr76^+/+; Apc^Min/+and Wdr76^−/−; Apc^Min/+ mice. Boxes indicate the enlarged areas. Tumors are indicated by dotted lines. T: Tumor. Scale bars represent 100 μm (left) and 20 μm (right). g Confocal immunofluorescence of Ki67 (red) and Ras (green) in tumor sections of Wdr76^+/+; Apc^Min/+and Wdr76^−/−; Apc^Min/+ mice. Tumors are indicated by dotted lines. Scale bars represent 20 μm