Fig. 4. Overexpression of lncRNA HEPFAL increases the sensitivity of erastin-induced ferroptosis.

A HCCLM3 and PLC cells transfected with lncRNA HEPFAL expression vector or control vector were treated with 20 μM erastin or untreated for 24 h and counted the surviving cells. B HCCLM3 and PLC cells were transfected with lncRNA HEPFAL expression vector and control vector treated with 20 μM erastin and were subjected to transmission electron microscopy. Representative images are shown. C, D Intracellular GSH levels were examined in HCCLM3 and PLC cells treated as (A). E, F FACS detected ROS levels in HCCLM3 and PLC cells treated as (A). G HCCLM3 cells stably overexpressing lncRNA HEPFAL were treated with 20 μM erastin to visualize the intracellular Fe²⁺ ions generation using the Fe²⁺ ions fluorescent probe Ferrorange. H Western blot analysis of PI3K, AKT, P-AKT, mTOR, SLC7A11, and GPX4 protein level in HCCLM3 and PLC cells transfected with lncRNA HEPFAL expression vector and control vector treated with 20 μM erastin. I, J Intracellular GSH levels were examined in HCCLM3 and PLC cells treated with DMSO, JR-AB2-011, and CCI-779. K, L FACS detected ROS levels in HCCLM3 and PLC cells treated as (I, J). *P < 0.05, **P < 0.01, ***P < 0.001.