p stat3 tyr705 rabbit polyclonal cell signaling technology  (Cell Signaling Technology Inc)

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Fe-capsaicin nanozyme attenuates sepsis-induced acute lung injury by regulating the functions of macrophages

doi: 10.3389/fbioe.2024.1509136

Figure Lengend Snippet: Antibodies used for Western blotting and immunofluorescence.

Article Snippet: P-STAT3 , Rabbit polyclonal , Cell Signaling Technology, United States , 9,145 , , 1:1,000.

Techniques: Western Blot, Immunofluorescence, Staining

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Fe-capsaicin nanozyme attenuates sepsis-induced acute lung injury by regulating the functions of macrophages

doi: 10.3389/fbioe.2024.1509136

Figure Lengend Snippet: Fe-CAP reduced the apoptosis of macrophages. Volcano plot (A) . GO enrichment analysis of Fe-CAP pretreated BMDM after LPS stimulation (top 20 pathways) (B) . KEGG enrichment analysis of Fe-CAP pretreated BMDM after LPS stimulation (top 18 pathways) (C) . Heat map analysis of Fe-CAP pretreated BMDM after LPS stimulation (D) . P-JAK2, JAK2, P-STAT3 and STAT3 levels were detected by Western blot (E) . The expression levels of Bcl-2/BAX/cleaved caspase-3 proteins in BMDM cells were evaluated by Western blot (E-1, E-2) .

Article Snippet: P-STAT3 , Rabbit polyclonal , Cell Signaling Technology, United States , 9,145 , , 1:1,000.

Techniques: Western Blot, Expressing

Journal: Journal of Inflammation Research

Article Title: Ginsenoside Rg1 Inhibits Microglia Pyroptosis Induced by Lipopolysaccharide Through Regulating STAT3 Signaling

doi: 10.2147/jir.s326888

Figure Lengend Snippet: Figure 3 Rg1 inhibits the phosphorylation of STAT3 induced by LPS in BV-2 microglia. (A) The target-PPI network of Rg1. As depicted, 17 crucial targets of Rg1 were identified. They are represented by the blue rounded square. In addition, the yellow ellipse represents Rg1. The red edge denotes the relationship between Rg1 and targets, while the the gray edge represents the PPI among Rg1’s targets. The network contains 18 nodes that are interconnected and associated with 41 edges. (B) Western blotting results showing phosphorylation of STAT3 in LPS- activated BV-2 microglial cells with or without Rg1 (20 and 60μM) pretreatment. Bar graph shows the gray value analysis based on immunoblot images. ***p < 0.001 compared with the control group, ###p < 0.001 compared with the LPS group. (C) Immunofluorescence images showing STAT3 expression (red) in lectin-positive microglia (green, arrows) in BV-2 microglia. DAPI-blue. Scale bars=20μm. (D) Note the enhanced expression of NLRP3 in activated microglia, which is noticeably attenuated by Rg1 treatment. *p < 0.05 compared with the control group, #p < 0.05 compared with the LPS group. (E) Immunofluorescence images showing STAT3 expression (red) in lectin-positive microglia (green, arrows) in the corpus callosum of postnatal brain. DAPI-blue. Scale bars=50μm. (F) Note enhanced expression of NLRP3 in activated microglia, which is obviously attenuated by Rg1 treatment. **p < 0.01 compared with the control group, #p < 0.05 compared with the LPS group.

Article Snippet: The rats in Rg1 Table 1 Antibodies Used for Immunofluorescence and Western Blotting Antibody Host Source Catalog Number Dilution for Staining Dilution for Western Blot NLRP3 Rabbit polyclonal Abclonal, China WH110563 1/200 1/1000 Cleaved-caspase-1 Rabbit polyclonal Cell Signaling Technology 4199s 1/1000 ASC Rabbit polyclonal Cell Signaling Technology 67824s 1/1000 β-Actin Mouse monoclonal Proteintech 66009-1-Ig 1/4000 STAT3 Mouse monoclonal Cell Signaling Technology 9139s 1/1000 p-STAT3 Rabbit polyclonal Cell Signaling Technology 9145s 1/100 1/1000 GSDMD Rabbit polyclonal Proteintech 20770-1-AP 1/2000 IL-1β Rabbit polyclonal Cell Signaling Technology 31202 1/1000 AIM Rabbit polyclonal Affinity DF3514 1/1000 Lectin Lycopersicon esculentum Sigma-Aldrich, USA L0401 1/200 DAPI Sigma-Aldrich, USA F6057 Mature-IL-1β Rabbit polyclonal Cell Signaling Technology 83186 1/1000 Pro-caspase-1 Rabbit polyclonal Cell Signaling Technology 24232 1/1000 GSDMD-N Rabbit polyclonal Cell Signaling Technology 36425 1/1000 Goat anti-rabbit IgG H&L (Alexa Fluor® 647) preadsorbed Goat anti-rabbit Abcam Ab150083 1/500 Horseradish peroxidase conjugated secondary antibody Goat anti-rabbit Affinity s0001 1/2000 Horseradish peroxidase conjugated secondary antibody Goat anti-mouse Affinity s0002 1/2000 https://doi.org/10.2147/JIR.S326888 DovePress Journal of Inflammation Research 2021:14 6622 R E T R C T E D Powered by TCPDF (www.tcpdf.org) treatment group were given Rg1 100mg/kg treatment 1h before LPS (1mg/kg) injection administrated intraperitoneally.

Techniques: Phospho-proteomics, Western Blot, Control, Immunofluorescence, Expressing

Journal: Journal of Inflammation Research

Article Title: Ginsenoside Rg1 Inhibits Microglia Pyroptosis Induced by Lipopolysaccharide Through Regulating STAT3 Signaling

doi: 10.2147/jir.s326888

Figure Lengend Snippet: Figure 4 Rg1 inhibits pyroptosis through STAT3 signaling. (A) Conserved sequences at binding sites of STAT3. (B) STAT3 binding sites on AIM2 promoters predicted by JASPAR. (C) The binding relationship between STAT3 and AIM2 promoter verified by ChIP-qPCR experiment. *p < 0.05 compared with IgG negative control. (D) Western blotting analysis showing NLRP3, ASC, AIM2, cleaved-caspase-1, pro-caspase-1, IL-1β, mature-IL-1β, GSDMD and GSDMD-N protein expression in BV-2 cells with or without Rg1 pretreatment exposed to LPS or LPS with IL-6 stimulation, and with or without stattic treatment induced by LPS. (E–M) Bar graph shows gray value analysis based on immunoblot images. *p < 0.05, **p < 0.01, ***p < 0.001 compared with control, #p < 0.05 and ###p < 0.001 compared with LPS treatment group, ∆p < 0.05 and ∆∆∆p < 0.001 compared with LPS+Rg1 treatment group. (N) Flow cytometry shows the percentage of dead cells over total cells in each stage. The effect of Rg1 (60μM) in down-regulating the rate of apoptotic cells induced by LPS was abrogated by IL-6. Compared with LPS (2μg/mL) treatment group, the rate of apoptotic cells was markedly decreased in stattic and LPS co-incubated group. (O) Bar graph shows the statistical results of the rate of dead cells in each group. **p < 0.01 compared with control, ##p < 0.01 compared with LPS treatment group, ∆∆p < 0.01 compared with LPS+Rg1 treatment group.

Article Snippet: The rats in Rg1 Table 1 Antibodies Used for Immunofluorescence and Western Blotting Antibody Host Source Catalog Number Dilution for Staining Dilution for Western Blot NLRP3 Rabbit polyclonal Abclonal, China WH110563 1/200 1/1000 Cleaved-caspase-1 Rabbit polyclonal Cell Signaling Technology 4199s 1/1000 ASC Rabbit polyclonal Cell Signaling Technology 67824s 1/1000 β-Actin Mouse monoclonal Proteintech 66009-1-Ig 1/4000 STAT3 Mouse monoclonal Cell Signaling Technology 9139s 1/1000 p-STAT3 Rabbit polyclonal Cell Signaling Technology 9145s 1/100 1/1000 GSDMD Rabbit polyclonal Proteintech 20770-1-AP 1/2000 IL-1β Rabbit polyclonal Cell Signaling Technology 31202 1/1000 AIM Rabbit polyclonal Affinity DF3514 1/1000 Lectin Lycopersicon esculentum Sigma-Aldrich, USA L0401 1/200 DAPI Sigma-Aldrich, USA F6057 Mature-IL-1β Rabbit polyclonal Cell Signaling Technology 83186 1/1000 Pro-caspase-1 Rabbit polyclonal Cell Signaling Technology 24232 1/1000 GSDMD-N Rabbit polyclonal Cell Signaling Technology 36425 1/1000 Goat anti-rabbit IgG H&L (Alexa Fluor® 647) preadsorbed Goat anti-rabbit Abcam Ab150083 1/500 Horseradish peroxidase conjugated secondary antibody Goat anti-rabbit Affinity s0001 1/2000 Horseradish peroxidase conjugated secondary antibody Goat anti-mouse Affinity s0002 1/2000 https://doi.org/10.2147/JIR.S326888 DovePress Journal of Inflammation Research 2021:14 6622 R E T R C T E D Powered by TCPDF (www.tcpdf.org) treatment group were given Rg1 100mg/kg treatment 1h before LPS (1mg/kg) injection administrated intraperitoneally.

Techniques: Binding Assay, ChIP-qPCR, Negative Control, Western Blot, Expressing, Control, Flow Cytometry, Incubation

Journal: Journal of Inflammation Research

Article Title: Ginsenoside Rg1 Inhibits Microglia Pyroptosis Induced by Lipopolysaccharide Through Regulating STAT3 Signaling

doi: 10.2147/jir.s326888

Figure Lengend Snippet: Figure 5 A schematic diagram depicting the proposed signaling mechanism via which Rg1 can exert its neuroprotection effect on lipopolysaccharide (LPS)-induced microglia pyroptosis. Following LPS stimulation, phosphorylated STAT3 (Tyr705) translocates into the nucleus and binds to the promoters of related genes such as AIM2 inflammasome. AIM2 transcription is initiated and forms inflammasome complexes with ASC and pro-caspase-1. Activated caspase-1 mediates the matura tion of IL-1β, and the cleaved N-terminal of GSDMD triggers pyroptosis. However, phosphorylation of STAT3 enhanced the expression of NLRP3. NLRP3, ASC and pro-caspase-1 are assembled into inflammasome complexes and mediate microglia pyroptosis. Rg1 disrupts the pathway and protects against LPS-induced microglia pyroptosis and inflammatory response through inhibition of STAT3 signaling.

Article Snippet: The rats in Rg1 Table 1 Antibodies Used for Immunofluorescence and Western Blotting Antibody Host Source Catalog Number Dilution for Staining Dilution for Western Blot NLRP3 Rabbit polyclonal Abclonal, China WH110563 1/200 1/1000 Cleaved-caspase-1 Rabbit polyclonal Cell Signaling Technology 4199s 1/1000 ASC Rabbit polyclonal Cell Signaling Technology 67824s 1/1000 β-Actin Mouse monoclonal Proteintech 66009-1-Ig 1/4000 STAT3 Mouse monoclonal Cell Signaling Technology 9139s 1/1000 p-STAT3 Rabbit polyclonal Cell Signaling Technology 9145s 1/100 1/1000 GSDMD Rabbit polyclonal Proteintech 20770-1-AP 1/2000 IL-1β Rabbit polyclonal Cell Signaling Technology 31202 1/1000 AIM Rabbit polyclonal Affinity DF3514 1/1000 Lectin Lycopersicon esculentum Sigma-Aldrich, USA L0401 1/200 DAPI Sigma-Aldrich, USA F6057 Mature-IL-1β Rabbit polyclonal Cell Signaling Technology 83186 1/1000 Pro-caspase-1 Rabbit polyclonal Cell Signaling Technology 24232 1/1000 GSDMD-N Rabbit polyclonal Cell Signaling Technology 36425 1/1000 Goat anti-rabbit IgG H&L (Alexa Fluor® 647) preadsorbed Goat anti-rabbit Abcam Ab150083 1/500 Horseradish peroxidase conjugated secondary antibody Goat anti-rabbit Affinity s0001 1/2000 Horseradish peroxidase conjugated secondary antibody Goat anti-mouse Affinity s0002 1/2000 https://doi.org/10.2147/JIR.S326888 DovePress Journal of Inflammation Research 2021:14 6622 R E T R C T E D Powered by TCPDF (www.tcpdf.org) treatment group were given Rg1 100mg/kg treatment 1h before LPS (1mg/kg) injection administrated intraperitoneally.

Techniques: Phospho-proteomics, Expressing, Inhibition