anti mouse cd45 pe cyanine7  (Thermo Fisher)

Journal: Neural Regeneration Research

Article Title: Hypoxia-preconditioned bone marrow–derived mesenchymal stem cells protect neurons from cardiac arrest–induced pyroptosis

doi: 10.4103/NRR.NRR-D-23-01922

Figure Lengend Snippet: HP-BMSCs suppress OGD-induced neuronal pyroptosis to some extent by inhibiting the MAPK and NF-κB pathways. (A) From left to right: BMSCs at passage 3 (P3), osteogenic differentiation BMSCs, and adipogenic differentiation BMSCs. The cells are typically spindle-shaped. Scale bars: 100 μm. (B) Flow cytometric analysis showing the expression of BMSC surface markers. The cells expressed CD29 and CD90 but did not express CD11b and CD45. (C) Immunofluorescence staining of primary cultured neurons for MAP2 (red, Alexa Fluor 594), whereas nuclei were labeled with DAPI (blue). Scale bars: 75 μm. (D) Western blotting analysis was performed to evaluate the expression of NLRP3, GSDMD-N, ASC, cleaved-caspase-1, NF-κB, JNK, and p38 MAPK in primary neurons from the indicated group ( n = 3). The results of parallel test were shown in Additional Figure 2 . (E–K) Quantitative analysis of NLRP3 (E), GSDMD-N (F), ASC (G), cleaved-caspase-1 (H), NF-κB (I), JNK (J), and p38 MAPK (K) protein expression levels as shown in D. Results are expressed as mean ± SD. * P < 0.05, vs . control group; # P < 0.05, vs . OGD group; & P < 0.05, vs . OGD + N-BMSCs group; $ P < 0.05, vs . OGD + HP-BMSCs group (one-way analysis of variance followed by Tukey’s multiple comparisons test). BMSCs: Bone marrow mesenchymal stem cells; HP-BMSCs: hypoxia preconditioned bone marrow mesenchymal stem cells; N-BMSCs: normal-cultured BMSCs; OGD: oxygen-glucose deprivation.

Article Snippet: BMSC-specific surface antigens were detected by flow cytometry (Lin et al., 2020) using CD29-FITC, CD90-PE, CD11b-PE, and CD45-FITC antibodies (BioLegend, San Diego, CA, USA).

Techniques: Expressing, Immunofluorescence, Staining, Cell Culture, Labeling, Western Blot, Control

Journal: Neural Regeneration Research

Article Title: Perfluoropentane-based oxygen-loaded nanodroplets reduce microglial activation through metabolic reprogramming

doi: 10.4103/NRR.NRR-D-23-01299

Figure Lengend Snippet: In vitro biological effects of PFP-OLNDs in BV2 microglia. BV2 microglia were pre-incubated with different concentrations of PFP-OLNDs for 3 hours and then exposed to LPS for 24 hours. (A, B) ROS formation detected by flow cytometry. (C) Representative histograms of CD45, MHC class II and CD86 expression on microglia of different groups. Unstained control (Con) is indicated in the shaded histogram. Blank represents the PBS-treated group. (D) Surface expression (MFI) of CD45, MHC class II and CD86 are shown. (E, F) Western blotting of IL-1β, IL-6, and iNOS. (G) ELISA of IL-1β, IL-6, and TNFα from BV2 cellular supernatant. Data are expressed as mean ± SEM ( n = 3). ** P < 0.01, *** P < 0.001, vs. LPS (one-way analysis of variance followed by Bonferroni post hoc test). DCFH-DA: 2′,7′-Dichlorodihydrofluorescein diacetate; ELISA: enzyme-linked immunosorbent assay; IL-1β: interleukin 1 beta; IL-6: interleukin-6; iNOS: inducible nitric oxide synthase; LPS: lipopolysaccharide; MFI: mean fluorescence intensity; MHC: major histocompatibility complex; PBS: phosphate buffered saline; PFP-OLNDs: perfluoropentane-based oxygen-loaded nanodroplets; ROS: reactive oxygen species; TNFα: tumor necrosis factor α.

Article Snippet: Surface staining was then performed using 5 μL of the following antibodies: anti-mouse CD45-PE-Cyanine7 (eBioscience, Cat# 25-0459-42, RRID: AB_1944375), anti-mouse major histocompatibility complex (MHC) class II (IA/I-E)-PE (eBioscience, Cat# 12-5321-82, AB_465928), and anti-mouse CD86 (B7-2)-PE-Cyanine5 (eBioscience, Cat# 15-0862-82, AB_468778) for 20 minutes on ice in the dark.

Techniques: In Vitro, Incubation, Flow Cytometry, Expressing, Control, Western Blot, Enzyme-linked Immunosorbent Assay, Fluorescence, Saline

Journal: Neural Regeneration Research

Article Title: Human-induced pluripotent stem cell–derived neural stem cell exosomes improve blood–brain barrier function after intracerebral hemorrhage by activating astrocytes via PI3K/AKT/MCP-1 axis

doi: 10.4103/NRR.NRR-D-23-01889

Figure Lengend Snippet: hiPSC–NSC–Exos suppress leukocyte infiltration on day 3 post-ICH. (A) Flow cytometry analysis of neutrophils (CD45 high CD11b + Ly6G + ), macrophages (CD45 high CD11b + F4/80 + ), CD4 + T cells (CD45 high CD3 + CD4 + ), CD8 + T cells (CD45 high CD3 + CD8 + ), NK cells (CD45 high CD3 – NK1.1 + ), and B cells (CD45 high CD3 – CD19 + ) that had infiltrated the brain tissue. (B) Quantification of leukocyte subsets that had infiltrated the brain tissue ( n = 7). (C) Microglial cell counts ( n = 7). (D) Flow cytometry analysis of neutrophils (CD11b + Ly6G + ), macrophages (CD11b + F4/80 + ), CD4 + T cells (CD3 + CD4 + ), CD8 + T cells (CD3 + CD8 + ), NK cells (CD3 – NK1.1 + ), and B cells (CD3 – CD19 + ) that had infiltrated the spleen. (E) Quantification of immune cell subsets that had infiltrated the mouse spleen ( n = 7). Data are presented as mean ± SEM. ** P < 0.01, *** P < 0.001 (Student’s t -test). All statistical values are presented in . Exo: Exosomes; hiPSC: human induced pluripotent stem cell; ICH: intracerebral hemorrhage; NK: natural killer; ns: not significant; NSC: neural stem cell; PBS: phosphate-buffered saline.

Article Snippet: The CD45 antibody was purchased from Invitrogen (Carlsbad, CA, USA), and all other antibodies were purchased from BioLegend (San Diego, CA, USA).

Techniques: Flow Cytometry, Saline

Journal: Neural Regeneration Research

Article Title: Human-induced pluripotent stem cell–derived neural stem cell exosomes improve blood–brain barrier function after intracerebral hemorrhage by activating astrocytes via PI3K/AKT/MCP-1 axis

doi: 10.4103/NRR.NRR-D-23-01889

Figure Lengend Snippet: Summary of statistical effect sizes in Figure 4

Article Snippet: The CD45 antibody was purchased from Invitrogen (Carlsbad, CA, USA), and all other antibodies were purchased from BioLegend (San Diego, CA, USA).

Techniques: Control