Fig. 2
From: NOTCH-mediated non-cell autonomous regulation of chromatin structure during senescence
NOTCH1 and JAG1 can non-autonomously repress SAHF formation in adjacent cells. a Schematic showing experimental set-up. IMR90 cells expressing doxycycline (DOX)-inducible N1ICD-FLAG were cultured with IMR90 ER:HRASG12V cells expressing mRFP with 100 nM 4OHT ± 1000 ng/mL DOX for 6 days. b Quantification of SAHF-positive red cells for the experiment outlined in a. Alone: mono-cultured IMR90 ER:HRASG12V cells; iN1ICD: DOX-inducible N1ICD-FLAG. c Representative images of co-cultures indicated (scale bar = 25 µm). Insets are unmerged DAPI images of the indicated cells (arrows). d Schematic showing experimental set-up. IMR90 ER:HRASG12V cells expressing mRFP were co-cultured with RPE1 cells stably expressing either mVenus or JAG1-mVenus for 6 days ±100 nM 4OHT. e Quantification of SAHF-positive red cells for the experiment outlined in d. f Representative images of co-cultures indicated (scale bar = 25 µm). Insets are unmerged DAPI images of the indicated cells (arrows). Note DAPI foci in RPE1 cells are not SAHFs. b, e Lines indicate the mean value of individual replicates. n = 3 biologically independent replicates for all conditions. Statistical significance calculated using one-way ANOVA with Tukey’s correction for multiple comparisons; ***p ≤ 0.001, NS = not significant
