Methamphetamine induces autophagy as a pro-survival response against apoptotic endothelial cell death through the Kappa opioid receptor

Ma, J; Wan, J; Meng, J; Banerjee, S; Ramakrishnan, S; Roy, S

Confocal and immunofluorescence microscopy. Immunofluorescence staining was carried out using antibodies against LC3 (Cell Signaling) and Beclin1 (Santa Cruz Biotechnology). DyLight TM488-conjugated donkey anti-rabbit IgG (Jackson Immuno Research Laboratories Inc.) was used as the secondary antibody. HUVECs were transiently transfected with LAMP1-GFP by using Lipofectamine 2000 (Life Technologies). LAMP1-GFP construct is a generous gift from Dr Sundaram Ramakrishnan (University of Minnesota, Minneapolis, MN, USA). Rhodaminephalloidin or DAPI (Life Technologies) was used to visualize F-actin and the nucleus, respectively. Fields were chosen randomly from various regions to ensure objectivity of sampling. Digital images at  400 magnification were acquired using a Nikon confocal microscope.