Fig 2: The prognostic value of CBX1-3 mRNA expression in ovarian cancer. Its Affymetrix ID is 201518_at, 226473_at and 200037_s_at. (A) OS curves are plotted for all ovarian cancer patients (n = 1,656). (B) PFS curves are plotted for all ovarian cancer patients (n = 1,435). (C) OS curves are plotted for all ovarian cancer patients (n = 655). (D) PFS curves are plotted for all ovarian cancer patients (n = 614). (E) OS curves are plotted for all ovarian cancer patients (n = 1,656). (F) PFS curves are plotted for all ovarian cancer patients (n = 1,435).

Fig 3: Activation of Wnt/ß-Catenin pathway by the interaction of CBX1 and HMGA2. A. Gene set enrichment analysis (GSEA) based on the TCGA data indicated that WNT signaling pathway was activated in HCC cases with CBX1 overexpression. B. Cells with CBX1 overexpression were treated with ß-Catenin siRNA or inhibitor XAV-939. Colony formation was performed to determine the effect of ß-Catenin inhibition on CBX1-mediated cell proliferation. C. Transwell assays were used to examine the cell migration. D. CoIP was performed using specific antibodies for CBX1 and HMGA2 in HCC cells. The colocalization of the two proteins was observed by confocal assays. E. Cells were transfected with HGMA2 overexpression vector and CBX1 overexpression vector or siRNA for 48 h. Western blot was used to examine the expression of ß-Catenin phosphorylation at Ser552, CBX1 and HMGA2.

Fig 4: Genomic binding and transactivation ability of L80A Oct4 are similar to those of WT Oct4 in ESCs, but L80A mutation interrupts the balanced Oct4 interactome.(A) Heatmaps of Oct4 ChIP-seq signals in WT and L80A Oct4 ESCs. (B) Representative Oct4 ChIP-seq tracks showing similar occupancy profiles of WT and L80A Oct4. (C) Oct4 ChIP-qPCR assay of WT and L80A Oct4 ESCs. Occupancy relative to a negative control region is shown. Error bars represent SD of biological triplicates. (D) The sox-oct motif is significantly enriched at the top 1000 WT and L80A Oct4 target sites. (E) The relative transcriptional activity levels of WT and L80A Oct4 were measured by luciferase reporter assay in WT/WT and L80A/L80A Oct4 ESCs. (F) Comparison of our Oct4 interactome with the three published Oct4 network studies. Overlapping proteins in different published datasets and novel proteins found in this study are listed in different color boxes. (G) Co-IP and Western blot (WB) showed decreased Oct4-Klf5 interaction and increased Oct4-Cbx1, Oct4-Ctr9, and Oct4-Cdc73 interactions in L80A Oct4 ESCs.

Fig 5: CBX1-promoted HCC cell proliferation and migration in vitro. A. The overexpression or knockdown of CBX1 protein in HCC cell lines was confirmed by western blot analysis. B. Cells were transfected with CBX1 overexpression vector or siRNAs on day 1 and day3. The related cell growth rates assessed by MTT assay were indicated. *P < .05, **P < .01. C. Colony formation assays were performed to demonstrate the effect of CBX1 on HCC cell growth. D. Transwell assays were used to show the cell migration in cells with CBX1 overexpression or silence. *P < .05, **P < .01.