Extended Data Fig. 1: Characterisation of live-cell transcription imaging in ESCs. | Nature Cell Biology

Extended Data Fig. 1: Characterisation of live-cell transcription imaging in ESCs.

From: The Polycomb system sustains promoters in a deep OFF state by limiting pre-initiation complex formation to counteract transcription

Extended Data Fig. 1: Characterisation of live-cell transcription imaging in ESCs.The alternative text for this image may have been generated using AI.

(a) Validation that the MS2x128 array is appropriately inserted into the first intron of the corresponding gene. Top: a schematic illustrating the PCR screening strategy. Bottom: PCR results for Zic2, E2f6, and Hspg2. The experiment was repeated twice. (b) Images of intronic RNA-FISH (red) and focalized MCP-GFP signal (green) indicating that MCP-GFP accumulates at sites where intronic RNA sequences for Zic2, E2f6, and Hspg2 are identified. Nuclei are labelled with 4′,6-diamidino-2-phenylindole (DAPI, blue) and outlined with a dashed line. Representative of at least 10 images each. (c) Genomic cChIP-seq snapshots for Zic2, E2f6, Meis1, HoxA7, HoxD locus (Polycomb genes) and Hspg2 (non-Polycomb gene) illustrating signal for RING1B-AID, H2AK119ub1, SUZ12 and H3K27me3. cnRNA-seq signal before and after 4h RING1B-AID depletion is also shown. The data used is from Dobrinic et al.21. (d) smRNA-FISH analysis of transcript-per-cell distributions for parental (MCP-GFP expressing) and MS2x128 array-containing cell lines. Source numerical data are available in source data.

Source data

Back to article page