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Fig. 1 | Epigenetics & Chromatin

Fig. 1

From: Pho dynamically interacts with Spt5 to facilitate transcriptional switches at the hsp70 locus

Fig. 1

Antagonistic binding patterns of HSF and PRC1 at the hsp70 locus. a Experimental scheme used throughout the study to measure the dynamics of mRNA/nascent RNA production, chromatin binding patterns of proteins and protein–protein interactions during the heat shock response. S2 DRSC cells were heat shocked at 37 °C for 15 min and thereafter allowed to recover from the heat shock at 25 °C for up to 90 min. At time points indicated in the scheme, cells were harvested and used for the following experimental procedures: mRNA quantification, ChIP-qPCR, co-IP and nascent RNA quantification. b Quantitative RT-PCR (qRT-PCR) detection of hsp70 transcript levels during the heat shock response detailed in a. Distance of the location of the primers used from the hsp70 transcription start site (TSS) is + 687 bp and has been depicted in the form of a cartoon below the data figure. The control line represents cells that were maintained at 25 °C for the entire duration of the time course detailed in a. cf ChIP-qPCR measurements of occupancy levels of RNA polymerase II CTD, S2P form of RNA polymerase II, heat shock binding factor (HSF) and Polyhomeotic (Ph, a component of PRC1) at the hsp70 locus during the heat shock response detailed in a. Distances of the location of the primers used from the hsp70 TSS are as follows: for c and f (+ 78 bp), for d (+ 802 bp) and for e (− 85 bp) has been depicted in the form of a cartoon below the data figure. The control line represents cells that were maintained at 25 °C for the entire duration of the time course detailed in a. Data information: In bf, data are presented as mean ± SEM for n = 2 (except for f where n = 3). Statistical significance was determined by performing a paired two-tailed t test, **P ≤ 0.01, *P ≤ 0.05 and ns = non-significant

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