Cy3 goat anti rabbit antibody

Cells transduced with shRR or shUSP18 lentiviral vectors were fixed with 4% paraformaldehyde for 20 min, washed three times with cold PBS and then permeabilized in 0.1% Triton X-100 for 15 min. After blocking in 5% goat serum for 1h, cells were incubated with 1:100 anti-STAT1 (CST, Danvers, MA) diluted in blocking solution at 4°C overnight. Cells were washed three times with PBS, and then Cy3 goat anti-rabbit antibody (Abcam, Cambridge, UK) was used as secondary antibody for 1h at room temperature. Nuclei was stained using 5μg/ml 4,6-diamidino-2-phenylindole (DAPI) for 10 min. Cells were washed three times with PBS, and then examined the distribution of STAT1 using fluorescence microscope (Carl Zeiss, Göttingen, Germany).

Sections of paraffin-embedded tissues were deparaffinized and underwent antigen retrieval. Sections were blocked with 1% bovine serum albumin, permeabilized with 0.1% Triton-X100 in PBS and incubated overnight with primary antibodies for cTnT (abcam), Pdk4 (proteintech), Vimentin (Cell Signaling Technology), Tpm1 (Invitrogen), CD31 (abcam), or Pparg (proteintech). These sections were subsequently stained with secondary antibodies, Alexa Fluor 488 goat anti-rabbit antibody (abcam), Cy3 goat anti-rabbit antibody (abcam); After sections were stained with DAPI (4’, 6’-diamidino-2-phenylindole), images were obtained by confocal microscope.