Fig 2: Dextran sulfate sodium increases connexin 43 and ten-eleven translocation-2 expressions in vivo. A: Left panel: Histograms show the normalized gene expression of connexin 43 (Cx43) in mice colon tissues, as detected by quantitative polymerase chain reaction (qPCR). Middle panel: Western blot of Cx43 protein in mouse colon tissues. Right panel: Densitometric analysis of protein expression after normalization to β-actin. Dextran sulfate sodium (DSS)-treated mice had the highest expression of Cx43 in their colons; B: Left panel: Histograms display the normalized gene expression of ten-eleven translocation-2 (TET-2) in mouse colon tissues, as detected by qPCR. Middle panel: Western blot of TET-2 protein expression in mouse colon tissues. Right panel: Densitometric analysis of protein expression after normalization to β-actin. DSS exposure resulted in enhanced TET-2 transcription and carbenoxolone inhibition of gap junctions significantly increased TET-2 expression; C: Immunofluorescence images of colon tissues stained for Cx43 with the mean fluorescence intensity (MFI) analysis. Scale bar 20 μm; D: Immunofluorescence images of mouse colon tissues stained for CD68 and TET-2. Histograms reflect the MFI analysis. High levels of CD68+ cells correlated with greater expression of Cx43 and TET-2 in DSS-exposed mice. Scale bar 10 μm. Experiments were repeated five times. One-way ANOVA, aP < 0.05; bP < 0.005; dP < 0.0001. MFI: Mean fluorescence intensity; CBX: Carbenoxolone; DSS: Dextran sulfate sodium.

Fig 3: Upregulation of MAPKAPK3, MAPK14, DUSP22, STAT1, and VCP in the bone marrow-derived monocytes/macrophages of old mice. (a) Schematic diagram of the protocol assessing the aging-related genes in monocyte/macrophage derived from young or aged mice. (b) Immunofluorescence staining of CD68 and cytokeratin in primary bone marrow-derived macrophages differentiated by RENCA cell conditioned media. Nuclei were visualized using DAPI. Scale bar = 100 μm. (c–g) qPCR analysis of MAPKAPK3, MAPK14, DUSP22, STAT1, and VCP gene expression in primary bone marrow-derived monocytes isolated from young (5-week-old) or old (72-week-old) mice. n = 3, Error = SE, * = p-value < 0.05, ** = p-value < 0.01 compared to young mice.

Fig 4: Co-localization of PDGF receptorβ (PDGFR-β) and of CD68-expressing cells in aortic plaques from high-cholesterol-fed rabbits.(A-E) Rabbit abdominal plaque sections stained with anti-PDGFR-β mAb (green color) and anti-CD68 (red color) mAb (Bars = 50 μm). Counterstained with DAPI.

Fig 5: Fluorescence microscopy images showing CD68 (microglia) and GFAP (astrocytes) immunoreactivity to the microdevice and microelectrode. (A) Coronal tissue slice at 2 weeks microdevice post-injection. (B) Axial tissue slice from rat #5 at 17 weeks microdevice post-injection. (C) Axial tissue slice from rat #5 at 17 weeks microelectrode post-implantation. Images I show CD68 (yellow), II show GFAP (green), and III merge I and II. The dashed red line shows the estimated location of the microdevice or microelectrode. The dashed blue line shows the injection track and outlines the dimensions of the injection tool.