Fig 1: Activated ABAC1/ApoA1 signaling is involved in the antidepressant action of c-MSST targeting the left V1 of CUMS mice(A and B) Heat map of the differentially expressed proteins analyzed by iTRAQ (Group A, Control mice; Group B, CUMS mice; Group C, Sham c-MSST mice; Group D, c-MSST mice), n = 3 mice per group.(C and D) PPI network of the differential proteins in the four groups. The red circles indicate the major/hub nodes of the PPI network.(E and F) Representative fluorescence images of the mouse V1. Immunofluorescent double labeling for ApoA1 (red)/NeuN (green) and ABCA1 (green)/NeuN (red). Nuclei were stained blue with DAPI. n = 4 mice per group. Scale bar, 20 µm.(G) Colocalization of the three neurocytemarkers with ApoA1 was quantified in the left V1 of control and CUMS mice. n = 8–9 mice per group. Data are expressed as the mean ±SEM. ***p < 0.001 versus Con-NeuN+/ApoA1+; ###p < 0.001 versus CUMS-NeuN+/ApoA1+ using ANOVA with Sidak correction.(H–J) Expression of ApoA1 and ABCA1 in the left V1 of control and CUMS mice. n = 6 mice per group. Data are expressed as mean ±SEM. *p < 0.05, **p < 0.01, ***p < 0.001 using ANOVA with Bonferroni correction. iTRAQ, isobaric tagging for relative and absolute quantification; PPI, protein-protein interaction; ApoA1, apolipoprotein A1; ABCA1, ATP-binding cassette, subfamily A, member one; NeuN, neuronal nuclei; DAPI, 4',6-diamidino-2-phenylindole; SPIO, superparamagnetic iron oxide; MF, magnetic field; c-MSST, combined magnetic stimulation system treatment; CUMS, chronic unpredictable mild stress; ns, non-significant. See also Figures S3 and S4.
Fig 2: Effect of RC extract on NeuN and SynI levels in LPS-induced anxiety mice. Representative immunohistochemistry of (A) NeuN and (B) SynI levels in the CA1, CA3, and DG area of control, model, and RC group mice; scale bar, 50 µm. Staining was quantified by mean IOD. Data are from 6 random images per section and n=4 mice per group. Mean±SD plotted (Control vs Model, ##p<0.01; Model vs RC, *p<0.05, **p<0.01).
Fig 3: Representative immunofluorescent inflammatory factor receptor TNF-αRII in TNC. Shown are NeuN (green) and inflammatory factor receptor TNF-αRII (red) double-labeled cells in the TNC (400×, Scale bar = 50 μm, n = 4, three sections/rat).
Fig 4: Representative immunofluorescent and protein expression of inflammatory factor receptor IL-1ßRI in TNC. Shown are NeuN (green) and inflammatory factor receptor IL-1ßRI (red) double-labeled cells in the TNC (400×, Scale bar = 50 µm, n = 4, three sections/rat).
Fig 5: Immunofluorescent analysis of GLUT2 and PC expression in CTX, HIPP, and BS in a mouse model of T1DM(A–C) 20 µm coronal sections of the mouse brain were stained with GLUT2 (green) and NeuN (red) and quantified in in the CTX (A), HIPP (B), and brainstem (BS) (C). Zoomed-in images are shown for the retro-splenial (RS) cortex, Cornu Ammonis 3 (CA3) of HIPP, and the DVC of BS.(D) Quantification of % positive pixels for GLUT2 using HALO software.(E–G) 20 µm coronal sections of the mouse brain were stained with pyruvate carboxylase (PC) (green) and NeuN (red) and quantified in in the CTX (E), HIPP (F), and BS (G). Zoomed-in images are shown for the RS, the CA3 of HIPP, and the DVC of BS.(H) Quantification of percentage of positive pixels for PC using HALO software.Values are presented as mean ± SEM (n = 4 biological replicates), *p < 0.05, **p < 0.01, ***p < 0.001, analyzed by Student’s t test. Scale bars are either 1 mm, 500 µm, or 100 µm.PC, pyruvate carboxylase; GLUT2, glucose transporter 2.
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