Fig 2: Bace-1 deletion induces microglial activation in the 5xFAD mouse model.(A) Representative three-dimensional (3D) reconstruction of confocal Z-stack images from the subiculum of WT and Bace-1–null mice. IBA-1–positive microglia processes are shown in red, surrounding the thioflavin S–positive amyloid plaques in green. Increased engulfment of amyloid is appreciable in Bace-1–deleted microglial cells compared to 5xFAD;Bace-1fl/fl;Cx3cr1CreER (−TAM). Scale bars, 20 μm. (B) Quantification of the number of plaque-associated microglia (N = 3 pairs of mice, 100 plaques randomly selected per genotype; ***P < 0.001, Student’s t test). (C and D) Imaris software–based quantification of microglial parameters expressed as averaged volume area and process lengths and represented as means ± SEM (N = 3, animals per genotype). Bace-1 deletion enhances microglial infiltration and cross-talk with Aβ plaques. (E) Representative 3D reconstruction of confocal Z-stack images from the subiculum of WT and Bace-1–null mice. CD68-positive immune cells are labeled in red, surrounding and overlapping with the thioflavin S–positive amyloid plaques in green. Increased colocalization of CD68, a lysosomal marker with amyloid plaques, is appreciable in Bace-1–deleted microglia cells compared to 5xFAD;Bace-1fl/fl;Cx3cr1CreER (−TAM). (F) Quantification of the percentage of CD68 region of interest (red channel) colocalized with thioflavin S–positive amyloid plaques (green channel) presented as means ± SEM. n = 25 plaques and surrounding microglia. **P < 0.01.

Fig 3: The comparison of M2 macrophage subpopulations count in glomeruli between stage II and stage III of membranous nephropathy. Quantitative analysis of CD68, M2a, M2b, and M2c in membranous nephropathy stage II and stage III, Significant correlation *p < 0.05.

Fig 4: Association of B3GNT3 expression with macrophage infiltration levels in PAAD. (A) PAAD tissue exhibiting B3GNT3+ but CD68− staining. (B) PAAD tissue exhibiting B3GNT3− but CD68+ staining. (C) Spearman regression analysis of B3GNT3 expression with macrophage maker CD68. P<0.05, cor=−0.606444. B3GNT3, UDP-GlcNAc:βGal β-1,3-N-acetylglucosaminyltransferase 3; cor, correlation; PAAD, pancreatic adenocarcinoma. 0, negative; 1, weak; 2, moderate; 3, strong.

Fig 5: OPN induces EndoMT(A) Immunohistofluorescent staining of CD68, α-SMA, and CD31 showing EndoMT-derived cells and neighboring macrophages in CRC tissues. Examples of EndoMT-derived cells and macrophages were indicated by arrows and *, respectively. (B) Immunohistofluorescent staining of OPN, α-SMA, and CD31 showing EndoMT-derived cells and OPN-expressing cells in CRC tissues. Examples of EndoMT-derived cells and OPN-expressing cells were indicated by arrows and stars, respectively. (C) mRNA levels of VE-cadherin, Tie1, Tie2, CD31, α-SMA, and fibronectin in HUVECs pre-incubated 16 h with 2% FBS-containing medium and then treated 15 h with PBS or 0.3 μg/ml of OPN. (D) Quantification of VE-cadherin, Tie1, Tie2, CD31, α-SMA, and fibronectin mRNA levels in HUVECs treated as described in (C). The mean ± SD values of 3 independent experiments are shown. *, P < 0.05 when compared with the data of PBS-treated cells. (E) VE-cadherin, Tie1, Tie2, CD31, α-SMA, and fibronectin levels in HUVECs pre-incubated 16 h with 2% FBS-containing medium and then treated 24 h with PBS or 0.3 μg/ml of OPN. (F) Gap-junction activities in HUVECs treated with PBS or OPN as described in (E). Representative dot plots from 3 independent Calcein-transfer assays are shown. The cells in the R2 regions were categorized as Calcein-accepting cells. The % of Calcein-accepting cells quantified by CellQuest software represent cellular gap-junction activities of the tested cells. (G) Cell migration activities in HUVECs treated with PBS or OPN as described in Materials and Methods. Cell migration was monitored for 15 h using time-lapse photography, and the movement tracks of 10 randomly selected PBS or OPN-treated HUVECs were analyzed by Image-Pro Plus software. Quantification of the accumulated migration distances is shown in the bottom panel. The data are the mean ± SD values of 3 independent experiments. *, P < 0.05 when compared with PBS-treated cells. (H) Invasiveness of PBS or OPN-treated HUVECs determined by Transwell invasion assay. The representative images shown were invasive cells on the filters of Transwell inserts. The mean ± SD values of 3 independent experiments are shown in the bottom panel. *, P < 0.05 when compared with PBS-treated cells.