Immunostaining of Cultured Cells

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).