Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Poliumoside alleviates microglia-mediated inflammation and blood-brain barrier disruption via modulating the polarization of microglia after ischemic stroke in mice.

doi: 10.1016/j.phymed.2025.156881

Figure Lengend Snippet: Fig. 3. Pol attenuated microglial activation in vivo after stroke. (A-B). Representative images of Iba1 and CD16/32 (A) and CD206 (B) labeled brain tissue on day 1 after stroke; (D) Microglial endpoints/cell; (E) Process length/cell of microglia; the ratio of CD16/32+ (C) and CD206+ (F) cells(n = 4–5); (G-I) Western blot bands of iNOS, CD86, Arg-1 and CD206 in the brain on day 1 after stroke and β-actin internal control-based optical density quantification(n = 4–5). Bar= 20 μm. Data are presented as means ± SD, were performed by One-way ANOVA followed by Tukey’s multiple comparisons test. ^p < 0.05, ^^p < 0.01, ^^^p < 0.001, ^^^^p < 0.0001 tMCAO vs sham group; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 Pol-M vs tMCAO group; #p < 0.05, ##p < 0.01,### p < 0.001, ####p < 0.0001 Pol-H vs tMCAO group; $p < 0.05, $$p < 0.01, $$$ p < 0.001, $$$$p < 0.0001 EDB vs tMCAO group, ns = no statistical difference.

Article Snippet: The primary antibody used in this study was: rabbit anti CD206 (1:1000, A02285–2, Boster Biological, China), rabbit anti Arg-1 (1:1000, A01106, Boster Biological, China), rabbit anti iNOS (1:1000, BA0326, Boster Biological, China), rabbit anti CD86 (1:1000, A00220–4, Boster Biological, China), rabbit anti-occludin antibody (1:1000 ab31721, abcam),rabbit anti Claudin 5 Monoclonal AntibodyAlexa FluorTM 488(1:100 Thermo Fisher Scientific),rabbit anti ZO-1 (1:1000,Proteintech,21,773–1 Wuhan, China),rabbit anti p-stat3 (1:2000, 9145T,CST),mouse anti Stat3 (1:1000,9139,CST),while the secondary antibody was either rabbit anti-mouse IgG (H + L) or rabbit anti-rabbit IgG (Abbkine, CA, USA).

Techniques: Activation Assay, In Vivo, Labeling, Western Blot, Control

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Poliumoside alleviates microglia-mediated inflammation and blood-brain barrier disruption via modulating the polarization of microglia after ischemic stroke in mice.

doi: 10.1016/j.phymed.2025.156881

Figure Lengend Snippet: Fig. 4. Pol regulates BV2 cell polarization in vitro. (A) Workflow of vitro experiment. (B) CCK8 assay was used to detect the toxicity of Pol on BV2 cells (n = 5); (C-D) The levels of IL-6 and TNFα mRNA in BV2 cells(n = 4), (E-G)) Levels of Inflammatory cytokine in BV2 cell culture (n = 4–5); (H-I) The levels of M1 type genes CD86, COX-2, (J-K) The levels of M2 type genes CD206, Arg-1 in BV2 cells (n = 4); (L-M) Immunofluorescence detection of BV2 cell polarization representative images and data visualization (n = 4); (N–O) Flow cytometry detection of BV2 cell polarization (n = 4). (P-Q) Apoptosis of neurons(n = 3). Bar= 100 μm. Data are presented as means ± SD, were performed by One-way ANOVA followed by Tukey’s multiple comparisons test. *p < 0.05, **p < 0.01, ****p < 0.0001; ns= no statistical ifference.

Article Snippet: The primary antibody used in this study was: rabbit anti CD206 (1:1000, A02285–2, Boster Biological, China), rabbit anti Arg-1 (1:1000, A01106, Boster Biological, China), rabbit anti iNOS (1:1000, BA0326, Boster Biological, China), rabbit anti CD86 (1:1000, A00220–4, Boster Biological, China), rabbit anti-occludin antibody (1:1000 ab31721, abcam),rabbit anti Claudin 5 Monoclonal AntibodyAlexa FluorTM 488(1:100 Thermo Fisher Scientific),rabbit anti ZO-1 (1:1000,Proteintech,21,773–1 Wuhan, China),rabbit anti p-stat3 (1:2000, 9145T,CST),mouse anti Stat3 (1:1000,9139,CST),while the secondary antibody was either rabbit anti-mouse IgG (H + L) or rabbit anti-rabbit IgG (Abbkine, CA, USA).

Techniques: In Vitro, CCK-8 Assay, Cell Culture, Immunofluorescence, Flow Cytometry

Journal: International Journal of General Medicine

Article Title: TRIM5 Promotes Systemic Lupus Erythematosus Through CD4(+) T Cells and Macrophage

doi: 10.2147/IJGM.S416493

Figure Lengend Snippet: Proportions of naive CD4(+) T cells and M2 macrophages in PBMCs of SLE and control groups. ( A ) The gating strategy for naive CD4(+) T cells and M2 macrophages in PBMCs. CD4(+) T cells: CD3+CD4+; naive CD4(+) T cells: CD3+CD4+CD45RA+CCR7+; macrophages: CD11B+CD68+; M2 macrophages: CD11B+CD68+CD206+. Representative flow cytometry plots show naïve CD4(+) T cells ( B ) and M2 macrophages ( C ) of PBMCs in HCs (n = 4) and SLE (n = 5). Absolute numbers of CD4(+) T cells, naive CD4(+) T cells ( D ), and macrophages, M2 macrophages ( E ). *P < 0.05.

Article Snippet: PBMCs were isolated from blood treated with the anticoagulant EDTA by layering over Ficoll lymphocyte fluid (17-1140-02 Ficoll PaqueTM PLUS, GE Healthcare, Sweden) and density gradient centrifugation at 400 × g. Two hundred microlitres of PBMC samples were then stained with a set of specific monoclonal antibodies (mAbs) (FITC Rat Anti-CD11b [M1/70, 561688, BD Pharmingen]; Alexa Fluor 647 Rat Anti-Human CCR7 [CD197, 3D12, 557734, BD Pharmingen]; PE-Cy7 Mouse Anti-Human CD45RA [HI100, 560675, BD Pharmingen]; PE Mouse Anti-Human CD206 [19.2, 555954, BD Pharmingen]; PerCP-Cy5.5 Mouse Anti-Human CD3[SP34-2, 552852, BD Pharmingen]; BV510 Mouse Anti-Human CD4[SK3, 562970, BD Pharmingen]) against surface markers and incubated at 4 °C for 30 minutes.

Techniques: Flow Cytometry

Journal: Materials Today Bio

Article Title: A novel tropoelastin-based resorbable surgical mesh for pelvic organ prolapse repair

doi: 10.1016/j.mtbio.2020.100081

Figure Lengend Snippet: Analysis of foreign body response to tropoelastin:PCL mesh implanted in an ovine vaginal surgery model of POP. Immunohistochemistry for (a) CD45 (b) HLA-DR (c) CD206 and (d) CD34 stained positive cells in brown in the epithelium and lamina propria of a tropoelastin:PCL explant, in comparison to (e–h) incision control ovine vaginal tissues. Immunofluorescence shows colocalization of CD45 + leukocytes (green) and CD206 + M2 macrophages (red; merge = yellow) at the (i) tropoelastin:PCL filament tissue interface. In tissue more distant to the filaments, CD45 + leukocytes (green in merge panel) were either M1 inflammatory or M0 uncommitted macrophages. (j) CD45 + leukocytes (green) colocalize with CD206 + M2 macrophages (red) in incision control ovine vaginal tissue. Representative images of n = 1 mesh implanted and n = 1 incision control ewe. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.).

Article Snippet: Immunohistochemistry staining was performed on FFPE sections following antigen retrieval using 0.1 M citrate buffer, blocking endogenous peroxidase with 3% H 2 O 2 , incubation with protein block (Dako) for 30 min at room temperature, using mouse anti-CD45 (0.5 μg/mL, BioRad) and mouse anti-CD206 (0.5 μg/mL, Dendritics) primary antibodies for 1 h at 37C.

Techniques: Immunohistochemistry, Staining, Immunofluorescence

Journal: Journal of Neuroinflammation

Article Title: Constitutively active microglial populations limit anorexia induced by the food contaminant deoxynivalenol

doi: 10.1186/s12974-022-02631-7

Figure Lengend Snippet: Immunohistochemistry conditions

Article Snippet: IBA-1/CD206 , Rabbit polyclonal anti-IBA1 (1:4000) 019-19741, Fujifilm Wako Goat polyclonal anti-CD206 (1:500) AF2535, R&D Systems , Donkey anti-Rabbit Alexa Fluor 594 (1:400) A21207, Invitrogen Donkey anti-Goat Alexa Fluor 488 (1:400) (A11055, Invitrogen) , Bovine serum albumine 3% Bovine serum albumine 3%.

Techniques: Immunohistochemistry, Blocking Assay, Plasmid Preparation