alexa fluor 488-labelled mouse anti-human cd206 mab  (R&D Systems)

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

Journal: Journal of Inflammation (London, England)

Article Title: Inflammatory M1-like macrophages polarized by NK-4 undergo enhanced phenotypic switching to an anti-inflammatory M2-like phenotype upon co-culture with apoptotic cells

doi: 10.1186/s12950-020-00267-z

Figure Lengend Snippet: NK-4 upregulates CD38, CD86, and CD206 expression on THP-1 macrophages. THP-1 macrophages were cultured with 4–5 μM NK-4 for 3 days. Cells were then fixed and stained for CD38 ( a ), CD86 ( b ), or CD206 ( c ) using specific antibodies (green). Nuclei were detected with Hoechst 33258 (blue). Results are representative of three independent experiments with similar results

Article Snippet: Cells were fixed with 4% paraformaldehyde in PBS for 15 min at room temperature, washed and permeabilized with 0.1% (v/v) Triton X-100 for 30 min. After overnight incubation in PBS containing 3% BSA at 4 °C, cells were treated with human FcR blocking reagent (Miltenyi Biotech, Auburn, CA) and were stained with the following antibodies: FITC-labelled mouse anti-human CD38 mAb (303504, BioLegend CNS, Inc., San Diego, CA), FITC-labelled mouse anti-human CD86 mAb (555657, BD PharMingen), Alexa Fluor 488-labelled mouse anti-human CD206 mAb (FAB25342G, R&D Systems).

Techniques: Expressing, Cell Culture, Staining