Journal: iScience
Article Title: CBX7C⋅PHC2 interaction facilitates PRC1 assembly and modulates its phase separation properties
doi: 10.1016/j.isci.2024.109548
Figure Lengend Snippet: The molecular interactions and functions of CBX7C in mESCs (A) The co-IP analyses of the interactions between CBX7C and the indicated PRC1 components in the mESCs cell line inducibly expressing CBX7C (mESC: iCBX7C-EGFP-Flag). The asterisks indicate the bands of RING1B and PCGF2. (B) Examination of the enrichment of the indicated proteins or histone modifications at the classical PRC1 target genes. ChIP-qPCR analyses of the enrichment of CBX7C-EGFP-Flag at the indicated known cPRC1 target genes and the effects on the enrichment of other PRC1 components or H2AK119ub at these loci in the stable mESC line. NC-1 ( Myc ) and NC-2 ( Klf4 ) were the confirmed negative controls in mESCs. The statistical graph represents the ChIP-qPCR results (significant differences between means were determined using Student’s t test: ns, not significant, p > 0.05; ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001). (C) RT-qPCR analyses of the expression of the selected PRC1 target genes are shown in (B). (D) Dual luciferase assays testing the effects of CBX7A, CBX7C, or PHC2 on the reporter expression. HEK293T cells stably expressing the Firefly luciferase were transfected with a plasmid expressing GAL4-DBD alone or fused with the indicated PRC1 component. The levels of the Renilla luciferase were used for data normalization. Data in (B–D) are represented as mean ± SD.
Article Snippet: Rabbit anti PCGF2 , Abclonal , Cat#A17327; RRID: AB_2770811.
Techniques: Co-Immunoprecipitation Assay, Expressing, Quantitative RT-PCR, Luciferase, Stable Transfection, Transfection, Plasmid Preparation