Fig 1: The effect of TEN and miR-668-3p on NSCs. (A) Western blot analysis showing the impact of Aβ1-42 induction, miR-668-3p inhibitor, and TEN on the protein levels of OXR1, p53, and p21. (B) CCK8 analysis showing the effect of Aβ1-42 induction, miR-668-3p inhibitor, and TEN on NSC proliferation. FCM analysis showing the effect of Aβ1-42 induction, miR-668-3p inhibitor, and TEN on NSC apoptosis (C), cell cycle (D) and ROS (E). ELISA analysis showing the impact of Aβ1-42 induction, miR-668-3p inhibitor, and TEN on the secretion of MDA (F), NO (G), SOD (H), and GSH (I) in NSCs. *, P<0.05, TEN group vs. Aβ1-42 group; #, P<0.05, TEN + inhibitor group vs. Inhibitor group. OXR1, oxidation resistance 1; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; Aβ1-42, amyloid β 1-42; TEN, tenovin-1; PI, propidium iodide; FITC, fluorescein isothiocyanate; G1, first gap of the cell cycle; S, synthesis of the cell cycle; G2, second gap of the cell cycle; ROS, reactive oxygen species; MDA, malondialdehyde; NO, nitric oxide; SOD, superoxide dismutase; GSH, glutathione peroxidase; NSCs, neuronal stem cells; CCK8, Cell Counting Kit-8; FCM, flow cytometry; ELISA, enzyme linked immunosorbent assay.
Fig 2: The interaction between miR-668-3p and OXR1 in NSCs in vitro. (A) IF analysis results of NSCs isolated from murine brain tissue. RT-qPCR analysis examining the impact of Aβ1-42 on miR-668-3p (B) and OXR1 (C) expression. RT-qPCR analysis results of the impact of Aβ1-42 on the expression of OXR1. (D) Western blot analysis result investigating the impact of Aβ1-42 on OXR1 protein expression. (E) RT-qPCR analysis indicating the effectiveness of the synthesized miR-668-3p mimics and inhibitor. (F) RT-qPCR analysis showing the effectiveness of the 3 constructed siRNAs targeting OXR1. (G) Dual luciferase analysis showing the binding of miR-668-3p and OXR1. (H) RT-qPCR analysis examining the impact of si-OXR1 on miR-668-3p expression. *, P<0.05, Inhibitor group vs. Inhibitor NC group; #, P<0.05, Mimic group vs. Mimic NC group. Aβ1-42, amyloid β 1-42; OXR1, oxidation resistance 1; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; IF, immunofluorescence; NC, negative control; si, small interfering; WT, wild-type; mut, mutant; NSCs, neuronal stem cells; RT-qPCR, quantitative reverse transcription polymerase chain reaction.
Fig 3: The effect of miR-668-3p and OXR1 on NSCs. (A) CCK8 analysis examined the impact of Aβ1-42 induction, miR-668-3p inhibitor, and si-OXR1 on NSC proliferation. FCM analysis examining the effect of Aβ1-42 induction, miR-668-3p inhibitor, and si-OXR1 on NSC apoptosis (B), cell cycle (C), and ROS (D). ELISA analysis examining the impact of Aβ1-42 induction, miR-668-3p inhibitor, and si-OXR1 on the secretion of MDA (E), NO (F), SOD (G), and GSH (H) of NSCs. #, P<0.05, Inhibitor + si-OXR1 group vs. Inhibitor + si-NC group. *, P<0.05. Aβ1-42, amyloid β 1-42; NC, negative control; si, small interfering; OXR1, oxidation resistance 1; PI, propidium iodide; FITC, fluorescein isothiocyanate; G1, first gap of the cell cycle; S, synthesis of the cell cycle; G2, second gap of the cell cycle; ROS, reactive oxygen species; MDA, malondialdehyde; NO, nitric oxide; SOD, superoxide dismutase; GSH, glutathione peroxidase; NSCs, neuronal stem cells; CCK8, Cell Counting Kit-8; FCM, flow cytometry; ELISA, enzyme linked immunosorbent assay.
Fig 4: Screening of mRNA and protein levels in AD. (A) The GSE150696 dataset, TargetScan, and starBase databases jointly analyzed downstream mRNA expression. (B) Heatmap showing OXR1 gene expression in GSE150696 dataset. (C) RT-qPCR analysis of OXR1 expression in AD mice. (D) IHC analysis showing OXR1 protein levels in AD mice. (E) IF analysis indicating the location and levels of OXR1 in AD mice. (F) Western blot analysis indicating protein levels of p53 and p21 in AD mice. *, P<0.05. OXR1, oxidation resistance 1; AD, Alzheimer’s disease; IHC, immunohistochemistry; IF, immunofluorescence; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.
Supplier Page from Abcam for Anti-OXR1 antibody