Journal: Journal of Neuroinflammation

Article Title: Presence and activation of pro-inflammatory macrophages are associated with CRYAB expression in vitro and after peripheral nerve injury

doi: 10.1186/s12974-021-02108-z

Figure Lengend Snippet: Secretion of cytokines by pro-inflammatory LPS/IFN-γ and anti-inflammatory IL-4/IL-13 -polarized macrophages following treatment with CRYAB. a mRNA levels of iNOS, IL-12p40, arginase-1, and IL-10 in LPS/IFN-γ- and IL-4/IL-13-polarized macrophages; representative of 3 experiments, n =3–4 wells/group. b – i Concentration of IL-6 ( b , f ), IL-1β ( c , g ), IL-12p40 ( d , h ), and TNF-α ( e , i ) secreted by pro-inflammatory LPS/IFN-γ ( b – e ) and anti-inflammatory IL-4/IL-13 ( f – i ) -polarized macrophages from WT (white bars) and Cryab −/− (black bars) animals grown for 48 h in the presence or absence of a CRYAB peptide; representative of 3 experiments, n =4 wells/group. Data represent mean ± sem, * p < 0.05 independent t -test

Article Snippet: The tissues were incubated overnight at 4 °C with rabbit anti-Iba1 (Wako Chemicals, 019-1974, 1:200) or goat anti-Iba1 (Novus Biologicals, NB100-1028, 1:100), rat anti-mouse CD16/32 (BD Biosciences, 553842, 1:500), goat anti-mouse CD206 (Santa Cruz Biotechnologies, sc-34577, 1:500), chicken anti-P0 (Abcam, ab39375, 1:500), rabbit anti-CD68 (Cell Signaling, 97778, 1:100), rabbit anti-arginase 1 (Cell Signaling, 93668, 1:50), rabbit anti-iNOS (BD Transduction Laboratories, 1:100; gift from Dr. Samuel David, McGill University), and DAPI (Invitrogen, D3571, 1:2000).

Techniques: Concentration Assay

Journal: Materials Today Bio

Article Title: Metformin glycyrrhetinic acid binary injectable hydrogel for synergistic tumor immunotherapy via spatiotemporal microenvironment remodeling

doi: 10.1016/j.mtbio.2025.102749

Figure Lengend Snippet: In vitro reprogramming of the tumor microenvironment (TME) by Fe 3 O 4 NPs@Met-GA-H. ( A, B ) Immunofluorescence analysis and quantitative analysis of HMGB1 exposure (A, red) and CRT exposure (B, red) in B16-F10 cells following different treatments. Blue: DAPI (n = 3). ( C ) Immunofluorescence staining of CD86,iNOS and CD206 in macrophages (n = 3). ( D ) Immunofluorescence staining of PD-L1 (n = 3). ( E-G )Quantitative analysis of CD86,iNOS and CD206 expression. ( H ) Quantitative analysis of PD-L1 expression (n = 3). Data are presented as Mean ± SD. Pairwise comparisons against the control group were performed using Student's t-test; differences among multiple groups were assessed by one-way ANOVA. Ns: not significant (P > 0.05); ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 versus Ctrl. # P < 0.05, ## P < 0.01, ### P < 0.001 versus the IL-4 group. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Alexa Fluor® 594 AffiniPure Goat Anti-Rabbit IgG (H + L) (Cat. No. 111-585-003, dilution: 1:250) was sourced from Jackson ImmunoResearch Inc. Rabbit anti-CD86 antibody (Cat. No. WL05184, dilution: 1:200), rabbit anti-iNOS antibody (Cat. No. WL0992a, dilution: 1:200), rabbit anti-CD206 antibody (Cat. No. WL06177, dilution: 1:200), and fluorescein isothiocyanate (FITC)-labeled goat anti-rabbit IgG (H + L) (Cat. No. WLA032, dilution: 1:200) were purchased from Wanlei Bio-Tech Co., Ltd. Rabbit anti-PD-L1 antibody (Cat. No. bs-1103R, dilution: 1:250) was obtained from Beijing Bioss Biotechnology Co., Ltd. PerCP/Cyanine5.5 anti-mouse CD45 Recombinant Antibody (Cat. No. 157612, dilution: 1:200), PE anti-mouse CD11c Antibody (Cat. No. 117308, dilution: 1:200), APC anti-mouse CD86 Antibody (Cat. No. 159216, dilution: 1:200), FITC anti-mouse CD80 Antibody (Cat. No. 104706, dilution: 1:200), PE/Cyanine7 anti-mouse I-A b Antibody (Cat. No. 116420, dilution: 1:200), Zombie R718TM Fixable Viability Kit (Cat. No. 423116, dilution: 1:200), FITC anti-mouse CD3 Antibody (Cat. No. 100204, dilution: 1:100), PE anti-mouse CD4 Antibody (Cat. No. 100408, dilution: 1:200), APC/Cyanine7 anti-mouse CD8a Antibody (Cat. No. 100714, dilution: 1:200), APC anti-mouse/human CD11b Antibody (Cat. No. 101212, dilution: 1:200), APC/FireTM 750 anti-mouse F4/80 Antibody (Cat. No. 123152, dilution: 1:200), PE/DazzleTM 594 anti-mouse CD206 (MMR) Antibody (Cat. No. 141732, dilution: 1:200), and PE/Cyanine7 anti-mouse CD86 Antibody (Cat. No. 159208, dilution: 1:200) were purchased from BioLegend.

Techniques: In Vitro, Immunofluorescence, Staining, Expressing, Control

Journal: PLoS ONE

Article Title: Role of Suppressor of Cytokine Signaling-1 In Murine Atherosclerosis

doi: 10.1371/journal.pone.0051608

Figure Lengend Snippet: Atherosclerotic plaques from Socs-1 −/− triple-KO (Socs-1 −/− ) mice showed an increased CD68 (red);iNOS (green) double-positive cell content. B. CD68 (red); CD206 (green) double-positive cells were hardly detected. C. Atherosclerotic plaques from Socs-1 −/− triple-KO (Socs-1 −/− ) mice contained slightly more MOMA-2 (red); Ly-6C (green) double-positive cells after 4 weeks of HCD as well as D. a higher number of Ly-6G positive cells (scale bar A–C: 20 µm, scale bar D:50 µm). N = 5–7 animals per group.

Article Snippet: In short, sections were incubated with rat anti-mouse CD68 (clone FA-11, AbD serotec, Oxford, UK) and rabbit anti-mouse iNOS (AbCam, Cambridge, UK) or rabbit anti-mouse CD206 (clone MR5D3, AbD serotec, Oxford, UK) or respective isotype controls followed by incubation with anti-rat Alexa-549 or anti-rabbit Alexa-488 (both Invitrogen, Molecular Probes, Darmstadt, Germany).

Techniques:

Journal: Scientific reports

Article Title: Microenvironment M1/M2 macrophages and tumoral progression vary within C57BL/6 mice from same substrain in prostate cancer model.

doi: 10.1038/s41598-024-65960-y

Figure Lengend Snippet: Figure 3. Quantification of M1 CD68 + iNOS + and M2 CD68 + CD206 + macrophages in murine prostate cancer tumor sections. (a) M1 DAPI + CD68 + iNOS + macrophage staining in prostate cancer tumor sections. (b) Comparison of total intratumoral M1 macrophages between mice from BL/6nA and BL/6nB nuclei *P = 0.0039. (c) Comparison of M1 macrophage populations between study regions for each group *P < 0.001. (d) M2 DAPI + CD68 + CD206 + macrophage staining in murine prostate cancer tumor sections (e) Comparison of total intratumoral M2 macrophages between mice from BL/6nA and BL/6nB nuclei. (f) Comparison of M2 macrophage subpopulations between study regions for each group *P < 0.001. Spleen. Macrophage staining control in spleen slice.

Article Snippet: For the M1 subpopulation, monoclonal rabbit anti-mouse iNOS antibody (NB300-605 Novusbio®) and donkey anti-rabbit IgG Alexa Fluor 647® antibody (Jackson Immunoresearch®) were used.

Techniques: Staining, Comparison, Control

Journal: Scientific reports

Article Title: Microenvironment M1/M2 macrophages and tumoral progression vary within C57BL/6 mice from same substrain in prostate cancer model.

doi: 10.1038/s41598-024-65960-y

Figure Lengend Snippet: Figure 4. M1, M2 and non-polarized macrophages population in the different tumor regions. (a) Representative image of the immunostaining for the detection of the M1, M2 phenotypes and non-polarized macrophages performed on the tumors obtained from the BL/6nA and BL/6nB groups. (b) Quantification of M1 phenotype macrophages (red), M2 (blue) and total macrophages (green) for each of the tumor regions studied in the BL/6nA group. (c) Quantification of M1 phenotype macrophages (red), M2 (blue) and total macrophages (green) for each of the tumor regions studied in the BL/6nB group. (d) Average percentage of M1 phenotype macrophages, CD68 + iNOS (red), M2 phenotype, CD68 + CD206 + (blue) and non-polarized macrophages CD68 + iNOS-CD206—(yellow) for each of the regions studied.

Article Snippet: For the M1 subpopulation, monoclonal rabbit anti-mouse iNOS antibody (NB300-605 Novusbio®) and donkey anti-rabbit IgG Alexa Fluor 647® antibody (Jackson Immunoresearch®) were used.

Techniques: Immunostaining