Fig. 1: RNA interactions regulate ATRX chromatin association.
From: Disruption of ATRX-RNA interactions uncovers roles in ATRX localization and PRC2 function
a Schematic for nuclear fractionation of MEFs with and without RNase A or RNase H treatment. b Western blot for ATRX, LSD1, and HNRNPC in MEF nuclear soluble and chromatin-bound fractions obtained with and without RNase A treatment as indicated on top. Representative blot from four independent experiments is shown. c Western blot for ATRX, LSD1, and HNRNPC in MEF nuclear soluble and chromatin-bound fractions obtained with and without RNase H treatment as indicated. Representative blot from three independent experiments is shown. d Top—Schematic of RNase A or RNase H treatment of MEFs for immunostaining. Bottom—Immunostaining of control (top) and RNase A or RNase H treated (bottom) MEFs with ATRX (red) and Cbx5 (red) antibodies. Percent of nuclei with pericentromeric Cbx5 or ATRX signal is shown along with total number of nuclei (n) quantified. Scale bar = 10 μm. e Immunostaining of control (left) and RNase A treated (right) MEFs with ATRX (red) and Npm1 (green) antibodies. Nucleus is stained with DAPI. Percent of nuclei with pericentromeric ATRX signal or nucleolar Npm1 signal is shown along with total number of nuclei (n) quantified. Scale bar = 10 μm. f Top—Western blot for ATRX and EZH2 in WT MEFs with and without Actinomycin D treatment. Bottom—RT-PCR analysis of minor satellite transcripts in WT MEFs with and without Actinomycin D treatment. Data are presented as mean values +/− SEM. P values are calculated using two-sided Student’s t test. g Immunostaining of WT MEFs with ATRX and Cbx5 antibodies (red) before and after Actinomycin D treatment. Percent of nuclei with pericentromeric Cbx5 or ATRX signal is shown along with total number of nuclei (n) quantified. Scale bar = 10 μm. h Cross-linking and immunoprecipitation in WT MEFs with IgG (gray) or ATRX antibodies (dark blue). qPCR analysis with minor satellite primers. Data are presented as mean values +/− SEM. P values are calculated using two-sided Student’s t test. Source data underlying Fig. 1b–h are provided as a Source Data file.
