Ds ri2 color ccd

Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was performed according to the manufacturer’s instructions (Roche #12156792910). In short, HUASMCs were plated in dishes (60 mm) with slides, and after AngII and/or ANGPT8 siRNA treatment, cells were embedded in paraffin for 30 min and then treated with 0.01% Triton for 2 min on ice. Slides were then treated with TUNEL reaction mixture in a humidified chamber and washed with PBS. Finally, slides were mounted using Vectashield mounting medium with DAPI (ZLI-9557, ZSGB-BIO, Beijing, China). Images were obtained with a Ni-UNikon Upright Microscope equipped with a DS-Ri2 color CCD (Nikon, Tokyo, Japan).

Immunohistochemical staining was performed as previously described [22 (link)] using primary antibodies against ANGPTL8 (1:200 dilution; Abcam, Cambridge, UK), α-smooth muscle actin (α-SMA) (1:200 dilution; ZM0003, ZSGB-BIO, Beijing, China), or galectin 2 (MAC-2) (1:200 dilution; Abcam, Cambridge, UK), followed by staining with secondary antibodies. Images were obtained with a Ni-UNikon Upright Microscope equipped with a DS-Ri2 color CCD (Nikon, Tokyo, Japan).