Fig. 6.

SLC38A5 enhances glutamine uptake and inhibits ferroptosis in osteosarcoma cells. (A) Comparison of glutamine uptake of the shSLC38A5 and shNC groups cells. (B-C) The OCR and ECAR of 143B cells in the shSLC38A5 and shNC groups were measured and quantified, ① oligomycin, ② FCCP, ③ Rotenone & Antimycin A, ④ glucose, ⑤ oligomycin, ⑥ 2-DG. (D-E) The effect of SLC38A5 knockdown on GSH and MDA levels in osteosarcoma cells. (F-I) The 143B cells were treated with glutamine (2 mM) and incubated for 48 h, and then the key proteins of the PI3K/AKT/mTOR signaling pathway were detected by western blot. (G-H) Western blot analysis of ferroptosis-related proteins including Nrf2 and GPX4. (J-M) Under treatment with varying concentrations of glutamine, western blot was performed to detect the protein levels of p-PI3K, PI3K, p-AKT, AKT, p-mTOR, and mTOR in two groups of 143B cells. Con: LV-Control group; SLC: LV-SLC38A5 group. The concentrations were as follows: Gln(-): 0 mM, Gln(+): 2 mM, Gln(++): 5 mM, and Gln(+++): 10 mM. Results from three independent experiments are summarized in a histogram format. Comparison between different concentrations: * P < 0.05, ** P < 0.01, *** P < 0.001, ns, not significant. Comparison between different groups: # P < 0.05, ## P < 0.01, ### P < 0.001, ns, not significant