Fig. 5
From: In-cell identification and measurement of RNA-protein interactions
RBM6 and ZZZ3 are required for XCI in vivo. a RNA immunoprecipitation (RIP) of the HA-tagged RBM6 protein. Left panel, western blot for RMB6. Right panel, RNA levels of the indicated transcripts in the input and in the immunoprecipitated eluates. All enrichments are normalized to GAPDH mRNA and to the input sample. Each RIP experiment was performed on two independent biological replicates. Data are presented as mean ± s.d.; unpaired t-tests: **P < 0.01. b RNA immunoprecipitation (RIP) of the HA-tagged ZZZ3 protein. Left panel, western blot for ZZZ3. Right panel, RNA levels of the indicated transcripts in the input and in the immunoprecipitated eluates. All enrichments are normalized to GAPDH mRNA and to the input sample as described in the ‘Methods’ section. Each RIP experiment was performed twice on independent biological replicates. Data are presented as mean ± s.d.; unpaired t-tests: **P < 0.01; *P < 0.05, not significant (ns). Full blots are provided as a Source Data file. c Representative RNA FISH images of Xist-induced cells upon depletion of the indicated proteins. Xist is shown in red and the X-linked gene Lamp2 in green. The dashed line delineates cell nuclei. Asterisks indicate Lamp2 expression from the active X chromosome. Arrowheads indicate Lamp2 expression from the inactive X chromosome that escapes XCI. Scale bars, 5 μm. d Quantification of cells with bi-allelic Lamp2 expression assessed with RNA FISH and expressed as fold ratio over RLuc control. Data from three independent experiments are represented as mean ± s.d.; Student’s t-tests: **P < 0.01; *P < 0.05; not significant (ns). Dashed line delineates the level of RLuc.
