Extended Data Fig. 3: Measurement of endogenous HSF1 concentration and HSE valency in the promoter of HSP genes.
(a) Schematic diagram showing the measurement process of HSF1 concentration. A calibration curve was obtained using a serial dilution of JF549 dye to get the relationship between the fluorescence intensity and dye concentration. Under the same imaging condition, JF549 labelled HaloTag knock-in HSF1 cells were imaged to obtain the mean fluorescence intensity of the nucleus. (b) Representative images of JF549 dye of different concentrations displayed using the 16 colours lookup table. 9 fields of view were captured to measure the average fluorescence intensity under different concentrations. (c) Representative image of HSF1 in HaloTag knock-in cells labelled using JF549 dye. The image was shown using the 16 colours lookup table. Images are representative of three independent experiments. (d) Calibration curve between JF549 concentration and fluorescence intensity. Data are displayed as mean ± s.d. n = 9 fields of view pooled from 3 independent experiments. The mean fluorescence intensity of HSF1 in HeLa cells was measured in 501 cells. The ribbon region represents the standard error of the linear fitting. (e) Pie plot showing the number of TTCnnGAAnnTTC motif in HSF1 Cut&Tag peaks. (f) Pie plot showing the number of GAAnnTTC motif in HSF1 Cut&Tag peaks.
