Ab167605

Isolated lung tissues were fixed in 4% paraformaldehyde, embedded in paraffin, cut into 4‐μm‐thick sections, and stained with haematoxylin and eosin. Infiltrating neutrophils were revealed by immunofluorescence using anti‐mouse Ly‐6G/Ly‐6C (Gr‐1) antibody (BioLegend) followed by Alexa 488‐conjugated‐goat anti‐rabbit IgG (Abcam) secondary antibody.
hMSCs were fixed with 4% paraformaldehyde for 15 min, then washed with PBS and permeated with 0.5% Triton X‐100 (V900502‐100ML, Sigma) in PBS for 15 min. The nuclei were stained with Hoechst 33324 (H3570, Thermo Fisher Scientific). The antibodies against Vimentin (ab92547, abcam), HP1α (ab109028, abcam), Lamin B1 (ab16048, abcam), Lamin A/C (ab8984, abcam), H3K9Me2/3(5327, CST), SP100 (ab167605, abcam), and PML (ab179466 and ab96051, abcam) were used as primary antibodies. Secondary antibodies were Alexa 488‐conjugated‐goat anti‐rabbit IgG (Abcam) and Alexa 555‐conjugated‐goat antimouse IgG (Thermo Fisher Scientific). Images were taken by a laser‐scanning confocal microscope (Leica TCS SP8, Leica, Germany).

Primary human monocyte-derived macrophages were polarized to “M1” phenotype on coverslips. The cells were fixed with 4% paraformaldehyde and then permeabilized in 0.2% Triton (Biorad)/PBS. Blocking buffer 2% BSA (Sigma-Aldrich) was added for 30 min. Rabbit polyclonal anti-SP140 antibody (dilution 1:200) (ab171141; Abcam) or mouse polyclonal anti-SP100 antibody (dilution 1:200) (ab167605; Abcam) were added for 2 h) followed by 2 h of secondary antibody, Polyclonal goat anti-Rabbit, Alexa Fluor546 (A-11035, Invitrogen) (dilution of 1:1000), or goat anti-mouse, Alexa Fluor546 (A-21123, Life Technologies) (dilution of 1:500), respectively. DAPI (Thermo Fisher) was used for nuclear detection.