Anti α sma mab

Cell immunostaining was performed as described previously [73 (link)]. Briefly, cells were fixed with 4% PFA and then blocked with 10% normal goat serum and 0.1% Triton X-100 in PBS. The cells were then incubated with primary antibodies overnight. On the next day, the cells were incubated with the secondary antibodies for 90 min and DPAI for 5 min. The cells were kept in PBS for observation. Images were taken with a Leica DMi8 fluorescence microscope. The following antibodies were used: anti-T mAb (1:100, Abcam, Catalog No. ab209665), anti-Sarcomeric α-Actinin mAb (1:100, Abcam, Catalog No. ab9465), anti-α-SMA mAb (1:100, Santa Cruz, Catalog No. sc32251), anti-Sox1 mAb (1:100, Abcam, Cat No. ab109290), goat anti-rabbit Alex Fluor 488-conjugated IgG (1:500, Invitrogen, Catalog No. A11008), goat anti-mouse Alexa Fluor Plus 555-conjugated IgG (1:500, Invitrogen, Catalog No. A32727).

Cultured cells were trypsinized into single cells and then briefly washed with PBS. Subsequently, the cells were fixed with 4% PFA for 15 min and incubated in 90% methanol overnight at −20 °C. The next day, the cells were re-suspended and incubated in 0.5% BSA diluted primary antibody for 1 h, with no antibody treatment as the control for flow cytometry gating. Next, the cells were incubated with secondary antibodies for 30 min. Finally, the cells were subjected to flow cytometry on a BD FACSCantoII Flow Cytometer (BD). The following antibodies were used: anti-T mAb (1:100, Abcam, Cat No. ab209665), anti-SOX1 mAb (1:100, Abcam, Cat No. ab109290), anti-Troponin T (cTnT) mAb (1:100, Invitrogen, Cat No. MA512960), anti-α-SMA mAb (1:100, Santa Cruz, Cat No. sc32251), goat anti-rabbit IgG (H+L) pAb (1:500, Invitrogen, Cat No. A-11008), goat anti-mouse IgG (H+L) pAb (1:500, Invitrogen, Cat No. A-11001).