Fig. 3
From: Inhibition of autophagy sensitizes cancer cells to Photofrin-based photodynamic therapy
Construction and characterization of HeLa cells with CRISPR/Cas9-mediated genomic KO of ATG5. a Schematic diagram showing ATG5 gene and the sgRNA target site at exon 6. Cas-9-mediated double-strand break near the PAM sequence are indicated by arrows. b HeLa-sgGFP and HeLa-sgATG5 cells were incubated for 24 h in the presence or absence of 10 μM chloroquine (CQ) to block lysosomal degradation. Whole-cell lysates were collected and ATG5 and LC3 expression was analyzed by Western blotting. c To confirm impaired autophagosome formation, HeLa-sgGFP and HeLa-sgATG5 cells were incubated as in (b) and subsequently stained with anti-LC3 antibody to visualize autophagosomes by immunofluorescence microscopy