Additional file 1 of Nanoparticulate MgH2 ameliorates anxiety/depression-like behaviors in a mouse model of multiple sclerosis by regulating microglial polarization and oxidative stress
Creators: name: Li, Zhenghao
name: Chen, Kefu
name: Shao, Qi
name: Lu, Hongtao
name: Zhang, Xin
name: Pu, Yingyan
name: Sun, Xuejun
name: He, Hua
name: Cao, Li
Publisher: name: figshare
Year: 2024
Subjects: subject: Medicine
subject: Neuroscience
subject: Pharmacology
subject: Chemical Sciences not elsewhere classified
subject: Immunology
subject: FOS: Clinical medicine
subject: Biological Sciences not elsewhere classified
subject: Science Policy
subject: Mental Health

description: Additional file 1: Figure S1. Confirming the concentration of MgH2 in vitro. Cell viability of BV2 in different concentrations of MgH2 (F = 16.26), n = at least 5 repeats per group. Data are presented as mean ± SEM. Figure S2. MgH2 treatment promotes the maturation of OPCs in EAE mice. (A) Representative anti-Ki67 (gray), anti-PDGFRα (red), and anti-Sox10 (green) immunofluorescence of the CC. (B) Quantitative analysis of PDGFRα+Sox10+ cells/mm2 (F = 0.4957) and Ki67+PDGFRα+Sox10+ cells/mm2 (F = 0.4059), n = 3 mice per group. (C) Representative anti-CC1 (red) and anti-Sox10 (green) immunofluorescence of the CC. (D) Quantitative analysis of CC1+Sox10+ cells/mm2 (F = 2.7514), n = 3 mice per group. Data are presented as mean ± SEM. Figure S3. Myelination and axons do not alter after 24-h restraint stress and MgH2 treatment. (A) Representative anti-MBP (red), anti-NF200 (green), and anti-SMI32 (purple) immunofluorescence of the CC. (B, C) Quantitative analysis of the MFI of MBP (F = 0.092) and NF200 (F = 0.0109), n = 3 mice per group. (D) Representative anti-Ki67 (gray), anti-PDGFRα (red), and anti-Sox10 (green) immunofluorescence of the CC. (E) Quantitative analysis of PDGFRα+Sox10+ cells/mm2 (F = 0.0323) and Ki67+PDGFRα+Sox10+ cells/mm2 (F = 0.1672), n = 3 mice per group. (F) Representative anti-CC1 (red) and anti-Sox10 (green) immunofluorescence of the CC. (G) Quantitative analysis of CC1+Sox10+ cells/mm2 (F = 0.1775), n = 3 mice per group. Data are presented as mean ± SEM. Figure S4. MgH2 treatment has no effect on the long-term proliferation of OPCs after 24-h restraint stress. (A) Schematic diagram displaying the cell proliferation test in 4 different groups. (B) Representative anti-BrdU (red), anti-Olig2 (green) and anti-Sox10 (purple) immunofluorescence of the CC. (C) Quantitative analysis of BrdU+Olig2+Sox10+ cells/mm2 (F = 0.1466), n = 3 mice per group. Data are presented as mean ± SEM. Figure S5. Number of infiltrating cells is reduced by MgH2 treatment after 24-h restraint stress. (A) Representative brain sections of H&amp;E staining in different regions of the hippocampus. (B) Quantitative analysis of the number of infiltrating cells in the hippocampus (F = 0.3478), n = 3 mice per group. (C) Representative brain sections of H&amp;E staining in the CC. (D) Quantitative analysis of the number of infiltrating cells in the CC (F = 2.1923), n = 3 vs. 3 mice per group. Data are presented as mean ± SEM. Figure S6. MgH2 treatment does not affect OPCs functions in vitro. (A) Schematic diagram displaying the study on functions of OPCs in vitro. (B) Representative anti-BrdU (red) and anti-Caspase3 (purple) immunofluorescence of OPCs in vitro at 48 h (in proliferation medium) and 72 h (in differentiation medium), respectively. (C, D) Quantitative analysis of the percentages of BrdU+ cells (F = 0.4577) and Caspase3+ cells (F = 2.2225) in (B), n = 1076 vs. 1014 cells (BrdU), n = 1145 vs. 1217 cells (Caspase3). (E) Representative anti-MBP (green) immunofluorescence of OPCs in vitro at 72 h (in differentiation medium). (F) Western blotting analysis relative MBP (F = 2.2225) and MAG (F = 47.8691) protein levels of OPCs in vitro at 72 h (in differentiation medium), n = 3 experiments per group. Data are presented as mean ± SEM.