XB-IMG-176847
Xenbase Image ID: 176847
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Figure 2. Nuclear Localization and Chromatin Association Defects of FHS SRCAP Protein.
(A) Schematic of differentiation of hESC to CNCCs.
(B) Biochemical fractionation of human CNCCs with immunoblotting against endogenous SRCAP protein (Kerafast antibody). 1×, 3×, and 9× protein dilutions. Cyto, cytoplasmic fraction; Sol.Nuc, soluble nuclear fraction; Chr-B, chromatin bound fraction. Predicted protein size on left.
(C) Confocal microscopy of anti-GFP immunofluorescence staining of CNCCs overexpressing FLAG-GFP tagged WT SRCAP, FHS mutant SRCAP, SRCAP AT hooks, and FLAG-GFP alone (blue, DAPI; red, tagged protein). Bottom panel shows merged image with DAPI staining. Scale bar, 10 μm.
(D) Quantification of cellular localization of overexpression proteins with nucleus defined by DAPI signal; 95% confidence intervals from quasibinomial glm model.
(E) Biochemical fractionation of CNCCs overexpressing FLAG-GFP tagged WT SRCAP, FHS mutant SRCAP, SRCAP AT hooks, FLAG-GFP alone with GFP antibody for SRCAP or control proteins, HSP90 to mark cytoplasmic fraction (Cyto), TFAP2A and pan-H2A.Z to mark chromatin-bound fraction (Chr-B). 1× and 3× dilution of each sample.
See also Figure S2. Image published in: Greenberg RS et al. (2019) Copyright © 2019. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View |