stat3 rabbit polyclonal ab  (Proteintech)

Journal: Advanced Science

Article Title: Deciphering the Role and Mechanism of Decidual Monocyte‐Derived Macrophage Infiltration in Obstetric Antiphospholipid Syndrome at Single‐Cell Resolution

doi: 10.1002/advs.202503480

Figure Lengend Snippet: Abnormal infiltration of monocyte‐derived CCR2 + macrophages in OAPS decidua and their pro‐inflammatory roles. A) Bar plot showing the proportions of MDMs in decidual immune cells from OAPS patients and HCs in scRNA‐seq data. B) FCM analysis of the proportion of CCR2 + macrophages in the decidua from OAPS patients (n = 11) compared with HCs (n = 18). C) Representative images for CCR2 + and CCR2 − macrophages from the decidua of OAPS patients and HCs under the transmission electron microscope. Scale bar: 2 µm. D) Bubble plot showing the functional gene expressions in different types of decidual myeloid cells based on scRNA‐seq data. E) Bar plot showing the GO terms of marker genes in decidual MDMs. F) Bar plot showing the KEGG terms of marker genes in decidual MDMs. G) GSEA indicates the term of inflammatory response is significantly enriched in decidual CCR2 + macrophages. H) Bar plot showing the relative expression levels of genes related to inflammatory factors in decidual CCR2 + and CCR2 − macrophages, as detected using RT‐qPCR (n = 3). I) Representative immunoblots and semi‐quantified results of Arginase 1, NF‐κB P65, and STAT3 expressions in decidual CCR2 + and CCR2 − macrophages from OAPS patients (n = 3). J) Representative immunofluorescence images for CD14 (green), CCR2 (red), IGFBP1 (croci), HLA‐G (cyan), and DAPI (blue) co‐staining in decidua from HCs. Scale bar: 1000 µm. K) Representative immunofluorescence images for CD14 (green), CCR2 (red), IGFBP1 (croci), HLA‐G (cyan), and DAPI (blue) co‐staining in decidua from OAPS patients. Scale bar: 2000 µm. L) Line chart showing the effects of medium supernatants from CCR2 + and CCR2 − macrophages on HTR‐8/SVneo proliferation, as measured by CCK‐8 (n = 3). M) Bar plot showing relative expression levels of CASP3 , MMP2 , and MMP9 genes in HTR‐8/SVneo treated with supernatants from CCR2 + and CCR2 − macrophages, as detected by RT‐qPCR (n = 3). Data in (A) and (B) are presented as mean ± SD and analyzed using Student's t ‐test. Data in (H) and (I) are presented as mean ± SD and analyzed using a paired t ‐test. Data in (L) and (M) are presented as mean ± SD and analyzed using one‐way ANOVA with Dunnett's multiple comparisons test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. OAPS, obstetric antiphospholipid syndrome; HCs, healthy controls; MDM, monocyte‐derived macrophage; FCM, flow cytometry; scRNA‐seq, single‐cell RNA sequencing; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; GSEA, Gene Set Enrichment Analysis; RT‐qPCR, real‐time quantitative polymerase chain reaction; SD, standard deviation; ANOVA, analysis of variance.

Article Snippet: The primary antibodies used: IκBα Mouse mAb (CST, USA, #4814, 1:1000), Phospho‐IκBα Rabbit mAb (CST, USA, #2859, 1:1000), Phospho‐NF‐κB p65 Rabbit mAb (CST, USA, #3033, 1:1000), NF‐κB p65 Rabbit mAb (CST, USA, #8242, 1:1000), IKKα Mouse mAb (CST, USA, #11930,1:1000), Phospho‐IKKα/β Rabbit mAb (CST, USA, #2697, 1:1000), GAPDH Mouse mAb (Proteintech, China, #60004‐1‐Ig, 1:5000), TLR4 Rabbit Polyclonal Ab (ABclonal, China, #A11226, 1:1000), Arginase‐1 Rabbit Polyclonal Ab (Proteintech, China, #16001‐1‐AP, 1:1000) and STAT3 Rabbit Polyclonal Ab (Proteintech, China, #10253‐2‐AP, 1:1000).

Techniques: Derivative Assay, Transmission Assay, Microscopy, Functional Assay, Marker, Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining, CCK-8 Assay, Flow Cytometry, RNA Sequencing, Real-time Polymerase Chain Reaction, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Hedgehog Signalling Modulates Immune Response and Protects against Experimental Autoimmune Encephalomyelitis

doi: 10.3390/ijms23063171

Figure Lengend Snippet: Histological and immuno-histochemical analyses of spinal cord cross-sections from ptch +/− and wt mice. MOG 35–55 -inoculated mice were sacrificed on day 20 after inoculation, and spinal cord sections were subjected to haematoxilin-eosin staining ( A , B ) or immuno-histochemical analysis with antibodies against CD11b ( C ), Iba-1 ( D ) and GFAP ( E ) or double staining for GFAP and Stat3 ( F ). Arrows in D point to microglia with typical activated or resting morphologies in wt or ptch +/− samples, respectively. Thick arrows in F indicate double-positive cells with anti-GFAP (brown cytoplasm) and anti-Stat3 (red nuclei), thin arrows show infiltrating immune cells and the asterisk marks polymorphic leukocytes. Magnitude scales are indicated on each micrograph.

Article Snippet: Primary antibodies used for immunohistochemistry analysis were CD11b rabbit monoclonal (Abcam, Cambridge, United Kingdom, Ab133357), GFAP rabbit monoclonal (MilliporeSigma, Burlington, MA, USA, 04-1062), Phospho Stat3 (Tyr705) rabbit polyclonal (Elabscience E-AB-20983), β-galactosidase rabbit polyclonal (Cappel, MP Biomedicals, Fountain Parkway Solon, Ohio, USA, 55976), NG2 rabbit polyclonal (Abcam ab101807), human/mouse, Olig1 mouse monoclonal (R&D systems, Minneapolis, MN, USA, MAB2417), Olig2 Rabbit monoclonal (EPR2673) and Iba-1 goat polyclonal (Abcam Ab5076).

Techniques: Staining, Double Staining