CD163 Antibody - #DF8235

Figure 7. Verification of PLAC8 expression and immune cell infiltration in verified cohort. (A) Representative results of protein expression levels of PLAC8, CXCL10 and 7 immune cell markers in IC/BPS tissue and UC tissue (Microscale: 100 μm). (B) The IHC score of PLAC8 in IC/BPS tissues and UC tissues. (C) The IHC score of CXCL10 in IC/BPS tissues and UC tissues. (D) Number of positive cells for seven immune cell marker in IC/BPS tissues and UC tissues. (E) The relative proportions of seven immune cell marker’s protein expression in IC/BPS tissues and UC tissues. IC/BPS, interstitial cystitis/painful bladder syndrome; UC, unaffected control; IHC, Immunohistochemistry. **P 

Fig. 1. Ruxolitinib inhibits the polarization of IL-4/13-induced fibrotic macrophages in RAW264.7 cells. RAW264.7 cells were exposed to Nintedanib (1 μM) or Ruxolitinib (5 μM and 10 μM) and IL-4/13 (20 ng/mL) for 12 h. (A) RT-qPCR was used to detect the mRNA expression levels of Spp1, Ym1, and CD206 (n = 3 per group). (B) Western blot analysis of Arg-1 and CD206 protein expression levels. (C) Immunofluorescence staining of CD206, CD163, and Arg-1 in RAW264.7 cells. (D) Semi-quantitative analysis of CD206, CD163, and Arg-1 immunofluorescence staining in RAW264.7 cells. (E) Western blot analysis of p-JAK1, p-JAK2, and p-STAT6 protein expression levels (n = 3 per group). (F) Quantification of p-JAK1/JAK1, p-JAK2/JAK2, and p-STAT6/STAT6 protein expression in RAW264.7 cells. Data are presented as mean ± SEM (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001.

FIGURE 2 Role of EMs exosomes on macrophage polarization. (A) The exosomes were labeled with a PKH26 fluorescent probe and then co-cultured with macrophages induced from THP-1 cells, and the uptake of exosomes by macrophages was observed using a fluorescence microscope (scale bar: 200 μm). (B) RT-qPCR was performed to detect the mRNA expression levels of macrophage polarization markers (iNOS and CD86 for M1 markers; CD206 and CD163 for M2 markers). (C) Western blot was used to detect the protein expression levels of macrophage polarization markers (iNOS and CD86 for M1 markers; CD206 and CD163 for M2 markers). (D) The proportion of CD86+ (M1) and CD206+ (M2) macrophages was detected using a flow cytometer. Data represent mean ± SD from three independent experiments. **: P < 0.01, ***: P < 0.001, compared with control exosomes.

Figure 7 Expression analysis of ASPM in KIRC and LIHC tissues. (A) Relative level of ASPM mRNA using quantitative RT-PCR; **p < 0.01. (B) The expression of ASPM was analyzed by Western-blot analysis using a compound samples. (C) Tumor infiltration of B cells, CD8+ T cells, and M2 macrophages in KIRC and LIHC using immunohistochemistry. 20 diagnosed cases of KIRC and 20 diagnosed cases of LIHC samples for immunohistochemistry. We validate the relationship between ASPM expression and B cells (marker: CD19), CD8+ T cells (marker: CD8A), and M2 macrophages (marker: CD163), we performed immunohistochemistry to assess ASPM, CD19, CD8A, and CD163. Muscle and lymph nodes as control samples. (a-d, f-i) Tumor infiltration of B cells, CD8+ T cells, and M2 macrophages in KIRC. (k-n, p-s) Tumor infiltration of B cells, CD8+ T cells, and M2 macrophages in LIHC. (a-d) High expression of ASPM (+++), CD8A (++), CD19 (++), and CD163 (++) in KIRC. (f-i) Low expression of ASPM (+), CD8A (+), CD19 (+), and CD163 (+) in KIRC. (k-n) High expression of ASPM (+++), CD8A (++), CD19 (++), and CD163 (++) in LIHC. (p-s) Low expression of ASPM (+), CD8A (+), CD19 (+), and CD163 (+) in LIHC. (e, o) lymph nodes was used as a positive control in KIRC (+++) and LIHC (+++). (j, t) muscle was used as a negative control in KIRC (-) and LIHC (-). The expression density of ASPM, CD8A, CD19, and CD163 in KIRC and LIHC tissues were quantitated by scoring staining intensity, including negative (–) and weak (+) staining, moderate (++) and strong (+ + +) staining, respectively.

FIGURE 5 | (A) Both THP-1 cells and THP-1 derived macrophages expressed PD-1 and CD68. T24 cells expressed PD-L1. (B) A co-IP assay showing the binding between CD68 and PD-1, hinting at the possibility of a CD68 and PD-1 interaction. (C) The binding of CD68 and PD-1 promoted THP-1 derived macrophages to M2 polarization whereas the blockage can reverse the process. (D) Molecular docking showing the interactions of CD68 and PD-1 through possible LAMP-like and IgV domains. SP indicates signal peptide and TM indicates transmembrane domain. (E) The cell viability assays of T24, THP-1 derived macrophages and PBMC co- culture experiments. T24 cell group and THP-1 derived macrophage group were the control groups. ***p < 0.001 (Student’s t-test).

Figure 5. The distribution of PRRSV receptors in immune organs was analyzed by immunofluorescent staining. The receptors of Sn and CD163 were analyzed in thymus, spleen, hilar lymph node, and mesenteric lymph node, respectively. Scale bars = 50 μm.