Fig 1: CBX2 is upregulated in GC cell lines and CBX2 knockdown inhibits the activation of the ß-catenin signaling pathway. (A) CBX2 expression in GC cell lines was analyzed using the Cancer Cell Line Encyclopedia. (B) Expression of CBX2 in normal or GC cells was determined via western blotting. (C) CBX2 mRNA levels were analyzed via RT-qPCR. (D) mRNA expression of CBX2 was also evaluated via RT-qPCR to determine the transfection efficiency of shRNA-CBX2. (E) Expression of ß-catenin, c-myc and cyclin D1 were determined via western blotting and (F) semi-quantified. Data are presented as the mean ± SEM from three independent experiments. *P<0.05, **P<0.01 and ***P<0.001 vs. GES-1 or Control. CBX2, chromobox homolog 2; GC, gastric cancer; RT-qPCR, reverse transcription-quantitative PCR; shRNA, short hairpin RNA; NC, negative control.
Fig 2: Upregulation of CBX2 induced glioma cell proliferation, invasion, and self-renewal. (A and B) CBX2 plasmid transfection efficiencies were verified by RT-PCR and western blotting (P < .01). (C and D) CCK-8 and colony formation assays demonstrated that CBX2 plasmid-mediated overexpression of CBX2 induced the proliferation of glioma cells (P < .01, P < .05). (E) Transwell assays showed that CBX2 upregulation induced invasion compared to that of the scramble group (P < .01). (F) The glioma stem cell frequency was increased following transfection with the CBX2 plasmid, and the effects were analyzed by sphere formation assays. (G) The upregulation of CBX2 reduced the level of E-cadherin and increased the expression of N-cadherin, slug, and snail.
Fig 3: Downregulation of CBX2 inhibited glioma cell proliferation, invasion, and self-renewal. (A) The data (UALCAN, http://ualcan.path.uab.edu/index.html) showed that the expression of CBX2 was higher than that in normal brain tissue (P < .001). (B) The level of CBX2 was higher in glioma tissues. And with the increase of glioma grade, the expression of CBX2 increased, the expression of CBX2 in Grade IV glioma tissues was the highest. (C and D) The expression of CBX2 was analyzed in glioma cell lines by RT-PCR and western blotting. (E and F) CBX2-shRNA transfection efficiencies were verified by RT-PCR and western blotting (P < .001). (G and H) CCK-8 and colony formation assays demonstrated that CBX2-shRNA-mediated knockdown of CBX2 decreased the proliferation of glioma cells (P < .01). (I) Transwell assays showed that CBX2 downregulation reduced invasion compared to that in the scramble group. (J) Sphere formation analysis showed that the downregulation of CBX2 decreased the glioma stem cell frequencies (P < .05). (K) Knockdown of CBX2 reduced the levels of N-cadherin, slug, and snail and increased the level of E-cadherin.
Fig 4: Knockdown of CBX2 repressed tumor growth in vivo. (A) Fluc bioluminescence images showing the tumors in the nude mouse xenograft model. (B) Overall survival analysis demonstrated that CBX2 downregulation improved the survival time (P < .05). (C) The mouse body weights were evaluated. (D) Images of HE-stained tumor tissue. (E) The expression of CBX2 and Ki67 was analyzed by IHC staining. (F and G) The expression of EMT markers, E-cadherin, N-cadherin, slug, and slug were detected with IHC.
Fig 5: YAP overexpression reverses the inhibitory effect of CBX2 knockdown on the ß-catenin signaling pathway. Expression levels of (A) YAP and (B) ß-catenin, c-myc and cyclin D1 were analyzed via western blotting. Data are presented as the mean ± SEM from three independent experiments. **P<0.01 and ***P<0.001 vs. Control; #P<0.05 and ##P<0.01 vs. shRNA-CBX2-1. YAP, yes-associated protein; CBX2, chromobox homolog 2; shRNA, short hairpin RNA; NC, negative control.
Supplier Page from Abcam for Anti-CBX2 antibody