Fig. 3: LIN28A changes interactors during ESC differentiation.
From: The RNA binding protein LIN28A mediates chromatin dynamics during neuronal differentiation
A Metascape-based enrichment analysis of the biological processes of LIN28A interactors identified in ESCs or SFEBs. B FunRich-based enrichment analysis of the cellular components of the LIN28A interactors identified in ESCs or SFEBs. C Venn diagram showing the large number of unique LIN28A interactors identified in ESCs and SFEBs, as well as the common proteins. D STRING analysis of proteins related to chromatin remodeling from LIN28A-interacting components uniquely identified in ESCs (medium-confidence, strength 0.53, False discovery rate 1.30 e-10). E Western blot analysis of LIN28A immunoprecipitates to assess its interaction with PRC2 core subunits (EZH2 and SUZ12) and accessory subunits (JARID2 for PRC2.2 and MTF2 for PRC2.1). Immunoprecipitation was performed using anti-FLAG beads in ESCs transfected with the LIN28A-FLAG expressing vector or the empty (Mock) vector. F, G Proximity ligation assay (PLA) to assess the interaction of LIN28A with PRC2. ESCs expressing LIN28A-FLAG were analyzed by PLA coupling the anti-FLAG antibody with the following antibodies: anti-P-SMAD1/5 (as negative control), anti-EZH2, anti-SUZ12 and anti-JARID2. Quantification of positive spots/cell (from >480 cells) from three independent (n = 3) biological replicates is shown in the graph (F). In the corresponding images, PLA positive spots are visible in green (G). Controls of single antibodies are shown in Supplementary Fig. 5F. Scale bars: 10 µm. Bars represent SD. Wilcoxon test: ***p ≤ 0.0001.
