Journal: Developmental dynamics : an official publication of the American Association of Anatomists

Article Title: Rodent monocyte-derived macrophages do not express CD163: Comparative analysis using macrophages from living boreoeutherians.

doi: 10.1002/dvdy.70036

Figure Lengend Snippet: FIGURE 9 Immunohistological comparison of CD163 and CD206 expression in various murine tumors. Immunohistochemical staining of CD163 and CD206 in MCA205, MC38, hepatoma, and histiocytic sarcoma. Scale bars: 200 μm.

Article Snippet: The membranes were cut and incubated with the following primary antibodies: anti-CD163 antibodies (clone EPR19518; Abcam) (for all animals), anti-CD204 antibodies (clone MCA1322GA; Bio-Rad) (for mouse), anti-CD206 antibodies (clone MCA2235GA; Bio-Rad) (for mouse), and anti-β-actin antibodies (clone C4; Santa Cruz Biotechnology, Dallas, TX, USA) (for all animals).

Techniques: Comparison, Expressing, Immunohistochemical staining, Staining

Journal: Developmental dynamics : an official publication of the American Association of Anatomists

Article Title: Rodent monocyte-derived macrophages do not express CD163: Comparative analysis using macrophages from living boreoeutherians.

doi: 10.1002/dvdy.70036

Figure Lengend Snippet: FIGURE 11 Expression of macrophage surface markers on recovered macrophages after clodronate liposome-mediated macrophage depletion. (A) CCL2 gene expression in the liver and spleen 3 days and 1 month post-clodronate administration in C57BL/6 mice. Data presented as mean ± SD. **, p < .01. (B) Western blot analyses of CD163, CD204, CD206 in the liver and spleen 1 month post-clodronate administration in C57BL/6 mice.

Article Snippet: The membranes were cut and incubated with the following primary antibodies: anti-CD163 antibodies (clone EPR19518; Abcam) (for all animals), anti-CD204 antibodies (clone MCA1322GA; Bio-Rad) (for mouse), anti-CD206 antibodies (clone MCA2235GA; Bio-Rad) (for mouse), and anti-β-actin antibodies (clone C4; Santa Cruz Biotechnology, Dallas, TX, USA) (for all animals).

Techniques: Expressing, Gene Expression, Western Blot

Journal: Developmental dynamics : an official publication of the American Association of Anatomists

Article Title: Rodent monocyte-derived macrophages do not express CD163: Comparative analysis using macrophages from living boreoeutherians.

doi: 10.1002/dvdy.70036

Figure Lengend Snippet: FIGURE 13 Expression of macrophage surface markers in the liver and spleen of obese mice. (A) Gene expression of CCL2, TNF-α, and IL-6 in the liver and spleen 3 months post-ND or HFD administration. Data presented as mean ± SD. *, p < .05. **, p < .01. (B) Western blot analyses of CD163, CD204, CD206 in the liver and spleen 3 months post-ND or HFD administration.

Article Snippet: The membranes were cut and incubated with the following primary antibodies: anti-CD163 antibodies (clone EPR19518; Abcam) (for all animals), anti-CD204 antibodies (clone MCA1322GA; Bio-Rad) (for mouse), anti-CD206 antibodies (clone MCA2235GA; Bio-Rad) (for mouse), and anti-β-actin antibodies (clone C4; Santa Cruz Biotechnology, Dallas, TX, USA) (for all animals).

Techniques: Expressing, Gene Expression, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Deficiency of Oncostatin M Receptor β (OSMRβ) Exacerbates High-fat Diet-induced Obesity and Related Metabolic Disorders in Mice *

doi: 10.1074/jbc.M113.542399

Figure Lengend Snippet: Adipose tissue inflammation in WT and OSMRβ−/− mice at 8 weeks on the HFD. The mice (8 weeks old) were fed an HFD for 8 weeks. A, total number of F4/80-positive cells per weights of adipose tissue in WT, OSMRβ−/−, and OSMRβ−/−(PF) mice (n = 6). EWAT, epididymal white adipose tissue. B and C, the percentages of CD11c-positive cells (B) and CD206-positive cells (C) among the F4/80-positive cells in WT, OSMRβ−/−, and OSMRβ−/−(PF) mice. D, the percentages of CD11c/CD206-double-negative cells among the F4/80-positive cells in WT, OSMRβ−/−, and OSMRβ−/−(PF) mice. E and F, the mRNA expression levels of inflammatory and anti-inflammatory markers (TNF-α, IL-1β, IFN-γ, CCR2, MCP-1, TLR4, IL-6, MGL1, MGL2, IL-10, IL-13, and adiponectin) in the adipose tissue (E) and SVF (F) of WT, OSMRβ−/−, and OSMRβ−/−(PF) mice (n = 6). ADPN, adiponectin. The data represent the mean ± S.E. *, p < 0.05; **, p < 0.01 WT versus OSMRβ−/− mice; #, p < 0.05 WT versus OSMRβ−/−(PF) mice, Student's t test.

Article Snippet: The cells were then incubated with the following primary antibodies at 4 °C for 30 min: fluorescein isothiocyanate-conjugated anti-F4/80 antibody (eBiosciences), phycoerythrin-conjugated anti-CD11c antibody (eBiosciences), and Alexa Fluor 647-conjugated anti-CD206 antibody (AbD Serotec).

Techniques: Expressing

Journal: The Journal of Biological Chemistry

Article Title: Deficiency of Oncostatin M Receptor β (OSMRβ) Exacerbates High-fat Diet-induced Obesity and Related Metabolic Disorders in Mice *

doi: 10.1074/jbc.M113.542399

Figure Lengend Snippet: Adipose tissue inflammation and glucose metabolism in WT and OSMRβ−/− mice at 2 and 4 weeks on the HFD. A, total number of F4/80-positive cells per weights of adipose tissue in WT and OSMRβ−/− mice (n = 6). B and C, the percentages of CD11c-positive cells (B) and CD206-positive cells (C) among the F4/80-positive cells in WT and OSMRβ−/− mice. D, the percentages of CD11c/CD206-double-negative cells among the F4/80-positive cells in WT and OSMRβ−/− mice. E and F, the mRNA expression levels of inflammatory and anti-inflammatory markers (TNF-α, IL-1β, IFN-γ, CCR2, MCP-1, TLR4, IL-6, MGL1, MGL2, IL-10, IL-13, and adiponectin) in the adipose tissue (E) and SVF (F) of WT and OSMRβ−/− mice (n = 6). G–N, the results of the ipGTTs (G and K) and ITTs (I and M) in WT and OSMRβ−/− mice at 2 weeks (G–J) and 4 weeks (K–N) on the HFD (n = 6). The AUC for blood glucose on the ipGTTs (H and L) and ITTs (J and N) was shown. ADPN, adiponectin. The data represent the mean ± S.E. *, p < 0.05; **, p < 0.01 WT versus OSMRβ−/− mice, Student's t test.

Article Snippet: The cells were then incubated with the following primary antibodies at 4 °C for 30 min: fluorescein isothiocyanate-conjugated anti-F4/80 antibody (eBiosciences), phycoerythrin-conjugated anti-CD11c antibody (eBiosciences), and Alexa Fluor 647-conjugated anti-CD206 antibody (AbD Serotec).

Techniques: Expressing

Journal: The Journal of Biological Chemistry

Article Title: Deficiency of Oncostatin M Receptor β (OSMRβ) Exacerbates High-fat Diet-induced Obesity and Related Metabolic Disorders in Mice *

doi: 10.1074/jbc.M113.542399

Figure Lengend Snippet: The effects of OSM on glucose metabolism, adipose tissue inflammation, and hepatic steatosis in ob/ob mice. ob/ob mice were injected intraperitoneally with either vehicle or recombinant mouse OSM (12.5 ng/g of body weight) twice a day for 7 days. A, the body weights in vehicle- and OSM-treated ob/ob mice (n = 6). B, the tissue weights in vehicle- and OSM-treated ob/ob mice (n = 6). EWAT, epididymal white adipose tissue; SWAT, subcutaneous white adipose tissue. C and D, blood glucose (C) and serum insulin (D) levels in vehicle- and OSM-treated ob/ob mice in the fasted states (n = 6). In the fasted states, mice were fasted overnight before the experiments. E–H, the results of the ipGTTs (E) and ITTs (G) in vehicle- and OSM-treated ob/ob mice (n = 6). The AUC for blood glucose on the ipGTTs (F) and ITTs (H) is shown. For ipGTTs, mice were fasted for 16 h and intraperitoneally injected with d-glucose (0.5 g/kg of body weight). For ITTs, mice were fasted for 4 h and intraperitoneally injected with insulin (5 unit/kg of body weight). I, total number of F4/80-positive cells per weights of adipose tissue in vehicle- and OSM-treated ob/ob mice (n = 6). J and K, the percentages of CD11c-positive cells (J) and CD206-positive cells (K) among the F4/80-positive cells in vehicle- and OSM-treated ob/ob mice. L, the mRNA expression levels of anti-inflammatory markers (IL-10, IL-13, MGL1, and MGL2) in the adipose tissue of vehicle- and OSM-treated ob/ob mice (n = 6). M, Oil Red O and PAS staining of the livers of vehicle- and OSM-treated ob/ob mice. CV, central vein. Scale bar = 100 μm. N and O, the content of triglycerides (N) and total cholesterol (O) in the livers of vehicle- and OSM-treated ob/ob mice in the fed state (n = 6). In the fed state, mice were fasted for 4 h before the experiments to eliminate the feeding effects on lipid metabolism. The data represent the mean ± S.E. *, p < 0.05; **, p < 0.01 vehicle versus OSM, Student's t test.

Article Snippet: The cells were then incubated with the following primary antibodies at 4 °C for 30 min: fluorescein isothiocyanate-conjugated anti-F4/80 antibody (eBiosciences), phycoerythrin-conjugated anti-CD11c antibody (eBiosciences), and Alexa Fluor 647-conjugated anti-CD206 antibody (AbD Serotec).

Techniques: Injection, Recombinant, Expressing, Staining

Journal: Stroke and vascular neurology

Article Title: Enhanced liver X receptor signalling reduces brain injury and promotes tissue regeneration following experimental intracerebral haemorrhage: roles of microglia/macrophages.

doi: 10.1136/svn-2023-002331

Figure Lengend Snippet: Figure 3 Enhanced LXR activation promotes M/M phenotype shifting. (A) Representative images of confocal tile scan showing whole day 7 lesion labelled with DAPI (blue), IL1β (green) and Iba1 (red). Scale bar, 200 µm. The size of images (note the longer scale bar in the GW3965 group) is adjusted to present the whole lesion within each frame. (B) Representative image of a three-dimensional (3D) reconstruction of a perihaematomal region in the vehicle group using Imaris. Scale bar, 5 µm. (C) Representative confocal images of perihaematomal region labelled with DAPI (blue), IL1β (green) and Iba1 (red), the left corner within dot lines orients the lesion centre. Scale bar, 100 µm. (D–F) Bar graphs comparing the Iba1+ percentage (D) IL1β+ percentage (E) Iba1+IL1β+ over Iba1+ proportions (F) between GW3965 treatment and DMSO control within the lesion region at day 7; the sample size was n=10 per group. (G) Representative images of confocal tile scan showing whole day 7 lesion labelled with DAPI (blue), CD68 (green), CD206 (red) and Arg1 (magenta). Scale bar, 200 µm. Scale bars are not equal. (H) Representative image of a 3D reconstruction of a perihaematomal region in the treatment group using Imaris. Scale bar, 10 µm. (I) Representative confocal images of perihaematomal region labelled with DAPI (blue), CD68 (green), CD206 (red) and Arg1 (magenta); the left corner within dotted lines denotes the lesion centre. Scale bar, 100 µm. (J–N) Bar graphs comparing the CD68+ percentage (J), CD206+ percentage (K), Arg1+ percentage (L) CD68+CD206+ over CD68+ proportions (M) and CD68+Arg1+ over CD68+ proportions (N) between GW3965 treatment and DMSO control within the lesion region at day 7; the sample size was n=10 per group. Significance is indicated as *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001; two-tailed, unpaired t-test (D, J–L) or two-tailed, unpaired t-test with Welch’s correction (E, F, M, N). Bar graphs show individual data points and represent mean±SD. LXR, liver X receptor; M/M, microglia/macrophage.

Article Snippet: After transferring to Hybond polyvinylidene difluoride (Amersham) membrane using a wet transfer system (Bio- Rad), membranes were blocked with 5% BSA and then probed overnight with the following primary antibodies: rabbit anti- human/mouse ABCA1 (1:1000; Cell Signaling, 96292), rabbit anti- mouse ApoE (1:1000; Cell Signaling, 49285), mouse anti- human/mouse IL- 1β (1:1000; Cell Signaling, 12242), rabbit anti- mouse iNOS (1:1000; Cell Signaling, 13120), rabbit anti- human/mouse CD206 (1:1000; Cell Signaling, 24595), rabbit anti- human/ mouse Arginase 1 (1:1000; Cell Signaling, 93668) and rabbit anti- mouse β-Actin (1:1000; Cell Signaling, 24595).

Techniques: Activation Assay, Control, Two Tailed Test