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. 2021 Jul 7;220(9):e202101092. doi: 10.1083/jcb.202101092

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© 2021 Crossley et al.

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Figure S5. — S9.6 immunostaining is sensitive to incubation temperature. (A) Representative confocal images of methanol-fixed HeLa cells. After fixation, coverslips were incubated in PBS and kept at 4°C or 37°C overnight, followed by S9.6 immunostaining. S9.6 signal is shown in green and DAPI in blue. (B) Quantification of mean nuclear S9.6 intensities for the conditions shown in A. (C) Same as in A but for GFP-dRNH1 staining. (D) Same as for B but for GFP-RNH1 staining. Scale bars are 10 microns. Box plots show median (box central line), 25% and 75% percentiles (box edges), and minimum and maximum values (whiskers). Data are combined from two biological replicates (n = 2), with at least 45 nuclei scored per condition per experiment. ***, P ≤ 0.001 (Mann-Whitney U test); ns, P > 0.05.