The Alexa-647 Click-iT™ Plus EdU Cell Proliferation Kit (Thermo
Fisher Scientific: C10640) was used to image host chromatin structure. Briefly,
NHDFs were cultured in cell culture media containing 10 μM EdU for 24 h;
the time at which labelling was performed is indicated in the relevant figure
legends. Click-iT reactions to detect Alexa-647-labeled DNA were performed
following manufacturers protocols. Then, direct stochastic optical
reconstruction microscopy (dSTORM) was used to collect super-resolution images
of chromatin structure, as previously described34 . Super-resolution microscopy was
performed using a Leica DMi8 S module Infinity TIRF microscope with Leica
DFC9000 sCMOS camera using a 100x oil immersion objective, and LAS X control
software (Leica Microsystems, Leica Application Suite X version 3.4.2.18368).
Imaging was performed in OxEA buffer (12% Sodium DL-lactate solution
(Sigma-Aldrich: L1375) with 50 mM cysteamine (Millipore-Sigma: 30070) and 1:100
dilution of OxyFluor (Sigma-Aldrich: SAE0059)) and data was processed using the
Thunderstorm (1.3–2014-11–08) plugin for Image-J35 .
Fisher Scientific: C10640) was used to image host chromatin structure. Briefly,
NHDFs were cultured in cell culture media containing 10 μM EdU for 24 h;
the time at which labelling was performed is indicated in the relevant figure
legends. Click-iT reactions to detect Alexa-647-labeled DNA were performed
following manufacturers protocols. Then, direct stochastic optical
reconstruction microscopy (dSTORM) was used to collect super-resolution images
of chromatin structure, as previously described34 . Super-resolution microscopy was
performed using a Leica DMi8 S module Infinity TIRF microscope with Leica
DFC9000 sCMOS camera using a 100x oil immersion objective, and LAS X control
software (Leica Microsystems, Leica Application Suite X version 3.4.2.18368).
Imaging was performed in OxEA buffer (12% Sodium DL-lactate solution
(Sigma-Aldrich: L1375) with 50 mM cysteamine (Millipore-Sigma: 30070) and 1:100
dilution of OxyFluor (Sigma-Aldrich: SAE0059)) and data was processed using the
Thunderstorm (1.3–2014-11–08) plugin for Image-J35 .