Journal: Veterinary Research

Article Title: Staphylococcus aureus mediates pyroptosis in bovine mammary epithelial cell via activation of NLRP3 inflammasome

doi: 10.1186/s13567-022-01027-y

Figure Lengend Snippet: NLRP3 inflammasome regulates ASC speck formation during S. aureus . Fluorescence microscopy images of MAC-T cells immunoassayed for ASC (red) with or without MCC950 after 4 h of S. aureus treatment.

Article Snippet: Caspase-1 inhibitor VX765 and NLRP3 inflammasome inhibitor MCC950 were obtained from Glpbio (USA) and used at final concentrations of 100 and 15 μM.

Techniques: Fluorescence, Microscopy

Journal: Veterinary Research

Article Title: Staphylococcus aureus mediates pyroptosis in bovine mammary epithelial cell via activation of NLRP3 inflammasome

doi: 10.1186/s13567-022-01027-y

Figure Lengend Snippet: NLRP3 inflammasome activation by S. aureus is essential for the generation of GSDMD-N and the release of IL-1β and IL-18. A Activated caspase-1 and B GSDMD-N or released C IL-1β and D IL-18 of MAC-T cells treated with or without NLRP3 inhibitor MCC950 or caspase-1 inhibitor VX765 for 1.5 h prior to treatment with S. aureus for 4 h. E Levels of IL-1β and F IL-18 released after S. aureus treatment of MAC-T cells for 4 h in the presence or absence of 25, 50, or 75 mM KCl.

Article Snippet: Caspase-1 inhibitor VX765 and NLRP3 inflammasome inhibitor MCC950 were obtained from Glpbio (USA) and used at final concentrations of 100 and 15 μM.

Techniques: Activation Assay

Journal: Cell Proliferation

Article Title: Aconitine induces cardiomyocyte damage by mitigating BNIP3‐dependent mitophagy and the TNFα‐NLRP3 signalling axis

doi: 10.1111/cpr.12701

Figure Lengend Snippet: A‐B, The viability of H9c2 cells incubated with aconitine alone or a combination of aconitine and Z‐VAD, MCC950 or MCC950 plus Z‐VAD; C‐D, Western blot analysis of NLRP3 and RIPK pathway‐related proteins in total lysates of H9c2 cells after the indicated treatments (* P < .05)

Article Snippet: The NLRP3 inhibitor MCC950, TNFα inhibitor Enbrel and aconitine were obtained from Shanghai Selleck Chemical Co., Ltd. (Shanghai, China) and Meilun Biotech.

Techniques: Incubation, Western Blot

Journal: Cell Proliferation

Article Title: Aconitine induces cardiomyocyte damage by mitigating BNIP3‐dependent mitophagy and the TNFα‐NLRP3 signalling axis

doi: 10.1111/cpr.12701

Figure Lengend Snippet: A, Western blot analysis of NLRP3 pathway activation after aconitine incubation for 24 h with or without the NLRP3 inhibitor MCC950; B, The subcellular location of NLRP3 and ASC used to detect NLRP3 inflammasome formation after aconitine incubation for 24 h with or without MCC950, scale bar: 6 μm

Article Snippet: The NLRP3 inhibitor MCC950, TNFα inhibitor Enbrel and aconitine were obtained from Shanghai Selleck Chemical Co., Ltd. (Shanghai, China) and Meilun Biotech.

Techniques: Western Blot, Activation Assay, Incubation

Journal: Cell Proliferation

Article Title: Aconitine induces cardiomyocyte damage by mitigating BNIP3‐dependent mitophagy and the TNFα‐NLRP3 signalling axis

doi: 10.1111/cpr.12701

Figure Lengend Snippet: A, Western blot analysis of NLRP3 pathway activation after aconitine incubation for 24 h with or without the TNFα inhibitor Enbrel or BNIP3 shRNA. B, Immunofluorescence analysis of BNIP3 and TOM20 to detect mitophagy after aconitine incubation for 24 h with or without the TNFα inhibitor Enbrel, scale bar: 10 μm

Article Snippet: The NLRP3 inhibitor MCC950, TNFα inhibitor Enbrel and aconitine were obtained from Shanghai Selleck Chemical Co., Ltd. (Shanghai, China) and Meilun Biotech.

Techniques: Western Blot, Activation Assay, Incubation, shRNA, Immunofluorescence

Journal: Cell Proliferation

Article Title: Aconitine induces cardiomyocyte damage by mitigating BNIP3‐dependent mitophagy and the TNFα‐NLRP3 signalling axis

doi: 10.1111/cpr.12701

Figure Lengend Snippet: A, Masson's trichrome staining of myocardial tissue sections of control, aconitine treated and aconitine plus BNIP3‐OE rats, scale bar: 1 mm. B, immunofluorescent staining of TUNEL and TNFα in myocardial tissue sections of control, aconitine treated and aconitine plus BNIP3‐OE rats, scale bar: 1 mm. C, The immunohistochemistry analysis of NLRP3, LC3, CTnI and CaMKII in myocardial tissue sections of control, aconitine treated and aconitine plus BNIP3‐OE‐treated rats, scale bar: 50 μm. * P < .05 vs control. ** P < .01 vs control. ## P < .01 to aconitine‐treated group

Article Snippet: The NLRP3 inhibitor MCC950, TNFα inhibitor Enbrel and aconitine were obtained from Shanghai Selleck Chemical Co., Ltd. (Shanghai, China) and Meilun Biotech.

Techniques: Staining, Control, TUNEL Assay, Immunohistochemistry

Journal: Journal of ginseng research

Article Title: Ginsenoside Rd protects cerebral endothelial cells from oxygen-glucose deprivation/reoxygenation induced pyroptosis via inhibiting SLC5A1 mediated sodium influx.

doi: 10.1016/j.jgr.2022.05.006

Figure Lengend Snippet: Fig. 2. Ginsenoside Rd suppressed OGD/R-activated NLRP3 inflammasome and pyroptosis in HBMECs. A-E. Representative blots (A) and quantitative analysis (BeE) of NLRP3, cleaved caspase-1, IL-1b and GSDMD-NT expression in HBMECs after OGD stimulation, with or without treatment of MCC950 (1 mM) or ginsenoside Rd (15 or 30 mM). F-G. Representative flow cytometric images (F) and quantitative data (G) of caspase-1þ/PI þ cell ratio were provided. Results were reported as mean ± SD (n ¼ 3). ***p < 0.001. n.s.: not significant.

Article Snippet: Ginsenoside Rd (purity: 98.27%), MCC950 (a selective NLRP3 inflammasome inhibitor, purity: 99.7%) and phlorizin (a competitive inhibitor of SLC5A1 and SLC5A2, purity: 99.83%) were purchased from Selleck (Houston, TX, USA).

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Andrade-Oliveira Salvianolic Acid B Modulates Caspase-1–Mediated Pyroptosis in Renal Ischemia-Reperfusion Injury via Nrf2 Pathway

doi: 10.3389/fphar.2020.541426

Figure Lengend Snippet: Salvianolic acid B (SalB) promotes Nrf2 nuclear activation and inhibits NLR family pyrin domain-containing 3 (NLRP3)/thioredoxin-interacting protein-thioredoxin1 (TXNIP) expression in ischemia-reperfusion (I/R) mice. (A–C) The expression of NLRP3 and TXNIP. (D) Immunofluorescence images (magnification ×200) showing the nuclear expression and localization of Nrf2 in the Sham, I/R, SalB-L, SalB-M, SalB-H groups. Blue: nuclear staining (DAPI); red: Nrf2; staining. Scale bar: 20 μm. (E) Representative western blots and (F–H) quantification of relative protein expression for nuclear Nrf2, keap1 and HO-1. (I–K) Superoxide dismutase (SOD), glutathione (GSH), and malondialdehyde (MDA) detected by a microplate reader. Data are represented as images or expressed as the mean ± SEM of each group from three separate experiments. *p < 0.05, **p < 0.01 vs. sham group; # p < 0.05, ## p < 0.01 vs. I/R group.

Article Snippet: After two–four passages, cells were cultured for 24 h and then were randomly divided into three groups: Control group: cells were cultured under normal conditions (5% CO2, 21% O2, 74% N2); Model group: the cells were incubated with different concentrations of SalB(purity>98%; 358153; Nanjing DASF Biotechnology Co.Ltd.) for 24 h; Positive control group: cells were incubated in certain concentration of NLRP3 inhibitor MCC950(S7809;Selleck), caspase-1 specific inhibitor VX-765(S2228;Selleck) for 2 h, and were then placed in a hypoxia incubator chamber (27310, stemcell) hypoxia (5% CO2, 95% N2) for 6 h and then reoxygenated for an additional 1 h (5% CO2, 21% O2, 74% N2).

Techniques: Activation Assay, Expressing, Immunofluorescence, Staining, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Andrade-Oliveira Salvianolic Acid B Modulates Caspase-1–Mediated Pyroptosis in Renal Ischemia-Reperfusion Injury via Nrf2 Pathway

doi: 10.3389/fphar.2020.541426

Figure Lengend Snippet: Nuclear factor erythroid-2 related factor 2 (Nrf2) nuclear expression is upregulated and NLR family pyrin domain-containing 3 (NLRP3)/thioredoxin-interacting protein-thioredoxin1 (TXNIP) is down-regulated after SalB treatment in H/R (A–C) NLRP3 and TXNIP expression were examined by western blot. (D) Immunofluorescence results (magnification ×400) showing the expression of Nrf2 under normal conditions (control), SalB treatment and H/R-treated HK-2 cells. Blue, nuclear staining (DAPI); red, Nrf2 staining. Scale bar: 20 μm. (E) Representative western blots and (F–H) quantification of relative protein expression for nuclear Nrf2, keap1 and HO-1. (I) Representative images of fluorescence of ROS probed by DCFH-DA. Data are represented as images or expressed as the mean ± SEM of each group from three separate experiments. **p < 0.01 vs. control group; # p < 0.05, ## p < 0.01 vs. H/R group.

Article Snippet: After two–four passages, cells were cultured for 24 h and then were randomly divided into three groups: Control group: cells were cultured under normal conditions (5% CO2, 21% O2, 74% N2); Model group: the cells were incubated with different concentrations of SalB(purity>98%; 358153; Nanjing DASF Biotechnology Co.Ltd.) for 24 h; Positive control group: cells were incubated in certain concentration of NLRP3 inhibitor MCC950(S7809;Selleck), caspase-1 specific inhibitor VX-765(S2228;Selleck) for 2 h, and were then placed in a hypoxia incubator chamber (27310, stemcell) hypoxia (5% CO2, 95% N2) for 6 h and then reoxygenated for an additional 1 h (5% CO2, 21% O2, 74% N2).

Techniques: Expressing, Western Blot, Immunofluorescence, Control, Staining, Fluorescence

Journal: Frontiers in Pharmacology

Article Title: Andrade-Oliveira Salvianolic Acid B Modulates Caspase-1–Mediated Pyroptosis in Renal Ischemia-Reperfusion Injury via Nrf2 Pathway

doi: 10.3389/fphar.2020.541426

Figure Lengend Snippet: The pyroptosis of acute kidney injury (AKI) signaling and protective effect of SalB through the nuclear factor erythroid-2 related factor 2 (Nrf2)/NLR family pyrin domain-containing 3 (NLRP3) pathway. I/R and H/R trigger pyroptotic cell death signaling, including upregulation of thioredoxin-interacting protein-thioredoxin1 (TXNIP) and suppression of Nrf2 expression, leads to NLRP3 oligomerization, ASC recruitment and subsequent caspase-1 activation. Fortunately, SalB treatment ameliorates AKI by regulating the Nrf2/NLRP3 pathway. The arrows represent promotion, while the inverted T represent inhibition. The effect of Salvianolic acid B (SalB) is shown in red.

Article Snippet: After two–four passages, cells were cultured for 24 h and then were randomly divided into three groups: Control group: cells were cultured under normal conditions (5% CO2, 21% O2, 74% N2); Model group: the cells were incubated with different concentrations of SalB(purity>98%; 358153; Nanjing DASF Biotechnology Co.Ltd.) for 24 h; Positive control group: cells were incubated in certain concentration of NLRP3 inhibitor MCC950(S7809;Selleck), caspase-1 specific inhibitor VX-765(S2228;Selleck) for 2 h, and were then placed in a hypoxia incubator chamber (27310, stemcell) hypoxia (5% CO2, 95% N2) for 6 h and then reoxygenated for an additional 1 h (5% CO2, 21% O2, 74% N2).

Techniques: Expressing, Activation Assay, Inhibition

Journal: Cell communication and signaling : CCS

Article Title: P2X7 receptor activation leads to NLRP3-independent IL-1β release by human macrophages.

doi: 10.1186/s12964-023-01356-1

Figure Lengend Snippet: Fig. 2 P2X7-mediated IL-1β release is NLRP3 independent. A THP-1 macrophages were primed with Pam3CSK4 and then stimulated with ATP, BzATP or nigericin for 3 h. Antagonists of P2X7 receptor (A-804598, oxATP), P2X4 receptor (5-BDBD) or P2X receptors (PPADS) were added 1 h before stimulation. IL-1β concentration in the supernatants was analyzed by ELISA. ATP, BzATP or nigericin induced IL-1β release was set to 100%. Mean + SEM (n = 3). One-sample t test against 100%, ns ≥ 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001. B THP-1 macrophages were primed with Pam3CSK4. Potassium chloride was added together with ATP, BzATP or nigericin for 3 h. IL-1β concentration in the supernatants was analyzed by ELISA. ATP, BzATP or nigericin induced IL-1β release was set to 100%. Mean + SEM (n = 3). One-sample t test against 100%, *P ≤ 0.05, ****P ≤ 0.0001. C THP-1 macrophages were primed with Pam3CSK4 and then stimulated with ATP, BzATP or nigericin for 3 h. NLRP3 inhibitors MCC950 or Bay 11–7082 were added 1 h before stimulation. IL-1β concentration in the supernatants was analyzed by ELISA. ATP, BzATP or nigericin induced IL-1β release was set to 100%. Mean + SEM (n = 3—4). One-sample t test against 100%, ns ≥ 0.05, *P ≤ 0.05, ***P ≤ 0.001, ****P ≤ 0.0001 (D) NLRP3 KO THP-1 macrophages were primed with Pam3CSK4. Cells were then stimulated with ATP, BzATP or nigericin for 3 h. IL-1β concentration in the supernatants was analyzed by ELISA. Mean + SEM (n = 3). Two-tailed two sample t test, ns ≥ 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001

Article Snippet: In selected experiments, cells were preincubated with the non-competitive P2X7 receptor antagonist A804598 (1 μM, 4473, Tocris Bioscience, Bristol United Kingdom), irreversible P2X7 receptor antagonist oxidized ATP (oxATP, 300 μM, 505758, Merck, Darmstadt, Germany), P2X4 receptor antagonist 5-BDBD (25 μM, SML0450-5MG, SigmaAldrich, Taufkirchen, Germany), P2X receptor antagonist PPADS (100 μM, 0625, Tocris Bioscience, Bristol, United Kingdom), NLRP3 inhibitor MCC950 (10 μM, 5479, Tocris Bioscience, Bristol, United Kingdom) or Bay 11–7082 (20 μM, B5556, Sigma-Aldrich, Taufkirchen, Germany), Caspase-1 inhibitor Ac-YVAD-cmk (40 μM, 10014, Biomol, Hamburg, Germany), pan-caspase inhibitor Z-VADfmk (40 μM, tlrl-vad, Invivogen, Toulouse, France), serine protease inhibitor AEBSF (300 μM, 50985.100, Biomol, Hamburg, Germany), cysteine protease inhibitor E64 (10 μM, 324890, Merck, Darmstadt, Germany), aspartic protease inhibitor pepstatin A (50 μM, 2936, Carl Roth, Karlsruhe, Germany) 1 h before stimulation.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Cell communication and signaling : CCS

Article Title: P2X7 receptor activation leads to NLRP3-independent IL-1β release by human macrophages.

doi: 10.1186/s12964-023-01356-1

Figure Lengend Snippet: Fig. 4 Influence of priming with different TLR ligands on IL-1β release. A THP-1 macrophages were primed with LPS or Pam2CSK4. Cells were then stimulated with ATP, BzATP or nigericin for 3 h. IL-1β concentration in the supernatants was analyzed by ELISA. Mean + SEM (n = 3). Two-tailed two-sample t test, ns ≥ 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001. B THP-1 macrophages were primed with Pam3CSK4, Pam2CSK4 or LPS. Gene expression of IL18 or IL1B was normalized to GAPDH. Mean + SEM (n = 3). One-way ANOVA followed by Tukey´s post-test, ns ≥ 0.05. C THP-1 macrophages were primed with Pam3CSK4, Pam2CSK4 or LPS and then stimulated with ATP, BzATP or nigericin. NLRP3 inhibitor (MCC950) was added 1 h before stimulation. IL-1β concentration in the supernatants was analyzed by ELISA. ATP, BzATP or nigericin induced IL-1β release was set to 100%. Mean + SEM (n = 3—5). One-way ANOVA followed by Tukey´s post-test, ns ≥ 0.05, *P ≤ 0.05, **P ≤ 0.01

Article Snippet: In selected experiments, cells were preincubated with the non-competitive P2X7 receptor antagonist A804598 (1 μM, 4473, Tocris Bioscience, Bristol United Kingdom), irreversible P2X7 receptor antagonist oxidized ATP (oxATP, 300 μM, 505758, Merck, Darmstadt, Germany), P2X4 receptor antagonist 5-BDBD (25 μM, SML0450-5MG, SigmaAldrich, Taufkirchen, Germany), P2X receptor antagonist PPADS (100 μM, 0625, Tocris Bioscience, Bristol, United Kingdom), NLRP3 inhibitor MCC950 (10 μM, 5479, Tocris Bioscience, Bristol, United Kingdom) or Bay 11–7082 (20 μM, B5556, Sigma-Aldrich, Taufkirchen, Germany), Caspase-1 inhibitor Ac-YVAD-cmk (40 μM, 10014, Biomol, Hamburg, Germany), pan-caspase inhibitor Z-VADfmk (40 μM, tlrl-vad, Invivogen, Toulouse, France), serine protease inhibitor AEBSF (300 μM, 50985.100, Biomol, Hamburg, Germany), cysteine protease inhibitor E64 (10 μM, 324890, Merck, Darmstadt, Germany), aspartic protease inhibitor pepstatin A (50 μM, 2936, Carl Roth, Karlsruhe, Germany) 1 h before stimulation.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Gene Expression

Journal: Cell communication and signaling : CCS

Article Title: P2X7 receptor activation leads to NLRP3-independent IL-1β release by human macrophages.

doi: 10.1186/s12964-023-01356-1

Figure Lengend Snippet: Fig. 5 Proposed mechanism for NLRP3-independent IL-1β release by human macrophages after P2X7 receptor activation. Activation of TLR2/1 by Pam3CSK4 leads to NF-κB mediated production of pro-IL-1β and components of NLRP3 inflammasome. Nigericin-mediated K+ efflux leads to NLRP3 inflammasome oligomerization causing serine protease- and caspase-dependent IL-1β release. In contrast, P2X7 receptor stimulation by BzATP potentially initiates two distinct IL-1β releasing mechanisms that are both NLRP3-independent. One mechanism relies on K+ efflux and requires activation of caspase-1 and serine proteases. The other mechanism is independent of K+ efflux, caspases, and serine-, cysteine-, and aspartic proteases, suggesting the involvement of other mechanisms for proteolytic processing of IL-1β

Article Snippet: In selected experiments, cells were preincubated with the non-competitive P2X7 receptor antagonist A804598 (1 μM, 4473, Tocris Bioscience, Bristol United Kingdom), irreversible P2X7 receptor antagonist oxidized ATP (oxATP, 300 μM, 505758, Merck, Darmstadt, Germany), P2X4 receptor antagonist 5-BDBD (25 μM, SML0450-5MG, SigmaAldrich, Taufkirchen, Germany), P2X receptor antagonist PPADS (100 μM, 0625, Tocris Bioscience, Bristol, United Kingdom), NLRP3 inhibitor MCC950 (10 μM, 5479, Tocris Bioscience, Bristol, United Kingdom) or Bay 11–7082 (20 μM, B5556, Sigma-Aldrich, Taufkirchen, Germany), Caspase-1 inhibitor Ac-YVAD-cmk (40 μM, 10014, Biomol, Hamburg, Germany), pan-caspase inhibitor Z-VADfmk (40 μM, tlrl-vad, Invivogen, Toulouse, France), serine protease inhibitor AEBSF (300 μM, 50985.100, Biomol, Hamburg, Germany), cysteine protease inhibitor E64 (10 μM, 324890, Merck, Darmstadt, Germany), aspartic protease inhibitor pepstatin A (50 μM, 2936, Carl Roth, Karlsruhe, Germany) 1 h before stimulation.

Techniques: Activation Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Induction of Autophagy by Pterostilbene Contributes to the Prevention of Renal Fibrosis via Attenuating NLRP3 Inflammasome Activation and Epithelial-Mesenchymal Transition

doi: 10.3389/fcell.2020.00436

Figure Lengend Snippet: PT and MCC950 attenuates TGF-β-induced NLRP3 inflammasome activation and EMT in NRK-52E cells. (A) Immunoblotting for NLRP3 inflammasome components NLRP3, ASC, caspase-1, degraded form of caspase-I, IL-1β, and degraded form of IL-1β in NRK-52E cells following treatment with DMSO (Control), PT 2 μM, TGF-β 2.5 ng/ml, or PT combined with TGF-β for 48 h (PT + TGF-β). (B) Immunoblotting for NLRP3, ASC, caspase-1, degraded form of caspase-1, and EMT markers fibronectin and α-SMA at 48 h following treatment with DMSO (Control), TGF-β 2.5 ng/ml, TGF-β combined with NLRP3 inhibitor MCC950 (MCC 10 nM), or MCC in NRK-52E cells. The membrane was probed with anti-GAPDH to confirm equal loading of proteins. Immunoblots are representative of at least three independent experiments.

Article Snippet: The NLRP3 inhibitor MCC950, UA assay kit and XOD fluorometric assay kit were purchased from Cayman Chemical Company (Michigan, United States).

Techniques: Activation Assay, Western Blot, Control, Membrane

Journal: Frontiers in Cell and Developmental Biology

Article Title: Induction of Autophagy by Pterostilbene Contributes to the Prevention of Renal Fibrosis via Attenuating NLRP3 Inflammasome Activation and Epithelial-Mesenchymal Transition

doi: 10.3389/fcell.2020.00436

Figure Lengend Snippet: Role of autophagy in attenuating NLRP3 activation and EMT in response to PT treatment. (A) Atg5 expression in NRK-52E cells stably transfected with Atg5 knockdown shRNA [Atg5-(1) and Atg5-(2)]. (B) The induction of autophagy by PT was measured by flow cytometry in NRK-52E and Atg5 knockdown NRK-Atg5-(2) cells. The data represent the means ± SD of three independent experiments; * P < 0.05 compared with Control groups; # P < 0.05 compared with NRK-52E cells. (C) The percentage of acridine orange positive staining cells in NRK and NRK-Atg5-(2) treated with PT (2.5 μM), TGF-β, or PT + TGF-β for 48 h. (D) The protein expression of autophagy markers (p62 and LC3-II), NLRP3 inflammasome components (NLRP3, caspase-1 and degraded form of caspase-1), and EMT markers (fibronectin and α-SMA) were detected by Western blotting in NRK and NRK-Atg5-(2) cells following 48 h treatment with TGF-β (TG) or PT combined with TGF-β (P + T). The membrane was probed with anti-GAPDH to confirm equal loading of proteins. Immunoblots are representative of at least three independent experiments. (E) Proposed model for renal protective effects of PT. PT significantly reduces UA production and XOD activities, prevents renal dysfunction, and ameliorates renal fibrosis in animal models. The mechanistic studies implicate that PT induces autophagy through AMPK activation to restrain TGF–triggered NLRP3 inflammasome activation and EMT, subsequently contributing to the protection of renal fibrosis.

Article Snippet: The NLRP3 inhibitor MCC950, UA assay kit and XOD fluorometric assay kit were purchased from Cayman Chemical Company (Michigan, United States).

Techniques: Activation Assay, Expressing, Stable Transfection, Transfection, Knockdown, shRNA, Flow Cytometry, Control, Staining, Western Blot, Membrane