Journal: Molecular Medicine

Article Title: CKS2 induces autophagy-mediated glutathione metabolic reprogramming to facilitate ferroptosis resistance in colon cancer

doi: 10.1186/s10020-024-00979-5

Figure Lengend Snippet: CKS2 mediates GSH metabolism reprogramming through autophagy activation in CC cells A , B WB measured expression levels of autophagy-related proteins (LC3I, LC3II, and p62); C IF detection assessed LC3B expression; D CCK-8 assay detected cell viability; E , F PI staining measured cell survival; G The kit for GSH and GSSG detection determined the level of GSH; H Glutamate colorimetric kit analyzed the level of glutamate; I Cysteine colorimetric kit assessed the level of cysteine; J Cystine uptake fluorescence kit determined cystine uptake level; K The kit for GSH and GSSG detected GSH/GSSG ratio; L , M BODIPY581/591 staining tested lipid ROS level; N , O DCFH-DA staining examined cell ROS level. Data are presented as the mean ± SD of three independent experiments. Statistical analysis was conducted by one-way ANOVA. * means P <0.05

Article Snippet: The slides were incubated overnight at 4 °C with LC3B antibody (ABclonal, China, A5618) and then with the secondary antibody conjugated with AF555 (Bioss, China, bs-0295G-AF555) at room temperature for 1 h. After the addition of DAPI (10 ug/ml) (Solarbio, Cat. No: C0065, China) for nuclear staining, the slides were incubated for 5 min, washed with PBS, sealed, and observed under an inverted fluorescence microscope.

Techniques: Activation Assay, Expressing, CCK-8 Assay, Staining, Fluorescence