Figure 4.

REST attenuates the effect of miR-494-3p on viability, ROS, ferroptosis, and neuron injury in erastin-induced LUHMES cells. Erastin-stimulated LUHMES were cotransfected with miR-494-3p inhibitor and sh-REST or REST-overexpressed plasmid and miR-494-3p mimics, respectively. (a and b) qRT-PCR showing the levels of miR-494-3p (a) and REST (b). (c) Cell viability was measured through the CCK-8 assay. (d) A Flow cytometer verified the change in ROS activity. (e) TEM images showing changes in mitochondrial morphology. Scale bar = 2 μm. Mitochondria are marked with blue arrows. (f and g) Immunofluorescence images showing ACSL4 (f) and TH (g) expressions. Magnification: 200×, scale bar = 100 μm. (h) Western blotting analysis showing REST, TH, NSE, ACSL4, and GPX4 levels. Results were representative data from triplicate experiments, and data are means ± SD. ^∗P < 0.05, ^∗∗P < 0.01, and ^∗∗∗P < 0.001 vs. inhibitor+sh-NC group; ^#P < 0.05, ^##P < 0.01, and ^###P < 0.001 vs. mimics+vector group.