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. 2017 Oct 6;8(54):92815–92826. doi: 10.18632/oncotarget.21590

Figure 5. BCAR4 activates Wnt/β-catenin signaling.

Figure 5

(A) BCAR4 interacted with β-catenin as shown by RNA pulldown. Biotin-labeled BCAR4 was incubated with HCT8 or SW480 cell lysates, followed by SDS-PAGE and Western blot. (B) β-catenin enriched BCAR4 in HCT8 and SW480 cells. Anti-β-catenin was added into HCT8 or SW480 cell lysates. Then enriched RNAs were eluted and extracted. RT-qPCR was used for analysis of BCAR enrichment. (C) β-catenin enriched BCAR4 in CC sample cells as shown by RNA IP. (D) BCAR4 was colocalized with β-catenin in CC sample cells as shown by RNA FISH. BCAR4 probe was biotin-labeled. Red, β-catenin; Green, BCAR4 probe; Nuclei were stained by DAPI. Scar bar, 10 μm. (E) BCAR4 overexpression prevented β-catenin from degradation in CC sample cells. V, oeVector; B, oeBCAR4. (F) BCAR4 knockdown promoted β-catenin degradation in CC sample cells. C, shCtrl; S, shBCAR4. (G) BCAR4 overexpression prevented β-catenin from degradation in HCT8 cells. After CHX addition, cells were collected at indicative time points and lysed, followed by SDS-PAGE and WB. GAPDH was chosen as loading control. (H) BCAR4 overexpression promoted β-catenin entry into nucleus and activation of the Wnt/β-catenin signaling pathway. V, oeVector; B, oeBCAR4. **P<0.01 by two-tailed Student's t test. All data presented are shown as means ± SD collected from three independent experiments.