Journal: Cell Death & Disease

Article Title: Inhibition of HtrA2 alleviated dextran sulfate sodium (DSS)-induced colitis by preventing necroptosis of intestinal epithelial cells

doi: 10.1038/s41419-019-1580-7

Figure Lengend Snippet: a Differentially expressed proteins in colons of control and DSS-treated mice. Three percent DSS was administered in drinking water to C57BL/6 mice for 7 days and replaced with fresh water thereafter. On day 10, colons were collected and protein levels were measured by quantitative proteomics. Each dot represents one protein. HtrA2 is indicated by red dot. X axis represents P value and Y axis represents fold change of colonic protein level between control and DSS-treated mice. n = 3 mice/group. b – d HtrA2 expression was decreased in colon of DSS-treated mice. Three percent DSS was used to induce colitis as described in a . On day 7 and day 10, colons were collected to analyze the protein levels of HtrA2, MLKL, phosphorylated MLKL, and GAPDH by immunoblotting with corresponding antibodies ( b ) or immunohistochemical (IHC) staining with anti-HtrA2 antibody ( d ). Densitometric analysis of immunoblotted proteins ( c ). Scale bar, 50 μm. e – g Three percent DSS was used to induce colitis as described in a , and UCF-101 (10-μm mol/kg mice) or DMSO was injected intraperitoneally every day for 10 days. Mice were killed on day 10 to measure the colon length ( e ); and body weight ( f ), and survival rate ( g ) was determined. In c , e , f , data are presented as means ± SEM. * P < 0.05; ** P < 0.01; *** P < 0.001 (two-tailed unpaired Student’s t test)

Article Snippet: HtrA2 shRNA (shHtrA2) and nontarget control shRNA (shNC) constructs were purchased from Cyagen Biosciences (China).

Techniques: Control, Quantitative Proteomics, Expressing, Western Blot, Immunohistochemical staining, Immunohistochemistry, Injection, Two Tailed Test

Journal: Cell Death & Disease

Article Title: Inhibition of HtrA2 alleviated dextran sulfate sodium (DSS)-induced colitis by preventing necroptosis of intestinal epithelial cells

doi: 10.1038/s41419-019-1580-7

Figure Lengend Snippet: a , b HT-29 cells were pretreated with Nec-1s (10 μM) or different doses of UCF-101 for 1 h, followed by stimulation with TNF-α (20 ng/mL)/Smac mimetic (2 μM)/Z-VAD (25 μM) for 8 h. PI positive cells were analyzed by PI/Hoechst staining ( a ), and cell viability was determined by CCK8 analysis ( b ). c – e HT-29 cells were stably infected with lentiviruses carrying scramble shRNA (shNC) or two different shRNAs targeting two individual sites of HtrA2 (shHtrA2-1 or shHtrA2-2). HtrA2 protein levels were detected by immunoblotting ( c ). Indicated cells were stimulated with T/S/Z for 8 h. PI positive cells were analyzed by PI/Hoechst staining ( d ), and cell viability was determined by CCK8 ( e ). In a , b , d , e , data are presented as means ± SEM. ** P < 0.01; *** P < 0.001 (two-tailed unpaired Student’s t test)

Article Snippet: HtrA2 shRNA (shHtrA2) and nontarget control shRNA (shNC) constructs were purchased from Cyagen Biosciences (China).

Techniques: Staining, Stable Transfection, Infection, shRNA, Western Blot, Two Tailed Test

Journal: Cell Death & Disease

Article Title: Inhibition of HtrA2 alleviated dextran sulfate sodium (DSS)-induced colitis by preventing necroptosis of intestinal epithelial cells

doi: 10.1038/s41419-019-1580-7

Figure Lengend Snippet: a HT-29 cells were pretreated with UCF-101 (50 μM) for 1 h, followed by stimulation with T/S/Z for different times as indicated. Phosphorylation of RIPK1, RIPK3, and MLKL, as well as their protein levels, were analyzed by immunoblotting with corresponding antibodies. b Silencing of HtrA2 inhibited formation of necrosome in T/S/Z treated HT-29 cells. HT-29-shNC or HT-29-shHtrA2 cells were stimulated with T/S/Z for indicated times. The association between RIPK1 and RIPK3 was analyzed by immunoprecipitation with anti-RIPK1 antibody, followed by immunoblotting. c HT-29 cells stably expressing HtrA2-3× Flag fusion protein were stimulated with T/S/Z for indicated times. The association between RIPK1 and HtrA2-3× Flag was analyzed by immunoprecipitation with anti-Flag antibody, followed by immunoblotting. d HtrA2 translocated from mitochondria to cytosol during necroptosis. HT-29 cells were stimulated with T/S/Z for different times as indicated. Thereafter, the cytosol and the mitochondrial fractions were subjected to SDS-PAGE. Immunoblotting was performed with specific antibodies for the indicated proteins. e HtrA2 translocated from mitochondria to cytosol from mitochondria to cytosol during DSS-induced colitis. The cytosol and the mitochondrial fractions of colon tissues from normal mice (day 0) and DSS-treated mice (day 7) were subjected to SDS-PAGE. Immunoblotting was performed with specific antibodies for the indicated proteins. In d, e, antibodies to TOM20 and GAPDH served as controls for purity of mitochondrial and cytosolic fractions, respectively

Article Snippet: HtrA2 shRNA (shHtrA2) and nontarget control shRNA (shNC) constructs were purchased from Cyagen Biosciences (China).

Techniques: Phospho-proteomics, Western Blot, Immunoprecipitation, Stable Transfection, Expressing, SDS Page

Journal: Scientific Reports

Article Title: Rotavirus Calcium Dysregulation Manifests as Dynamic Calcium Signaling in the Cytoplasm and Endoplasmic Reticulum

doi: 10.1038/s41598-019-46856-8

Figure Lengend Snippet: RV-induced dynamic Ca 2+ signaling is related to virus dose. ( A) Immunofluorescence images of mock or SA114F-infected MA104-GCaMP5G cells that were inoculated with increasing MOI (0.1, 1, or 10). RV antigen (red) is detected at ~8 hpi with anti-RV polyclonal antisera and nuclei are stained with DAPI (blue). (B , C) Epifluorescence images of MA104-GCaMP5G cells mock or infected with recombinant SA11-mRuby reporter virus with increasing MOIs (0.1, 1, 10). Images were captured at 7 HPI (B) or 10 HPI (C) . (D) Representative single-cell traces of relative GCaMP5G fluorescence (F/F 0 ) from cells mock (black) or RV infected by SA114F with MOIs of 0.1 (purple), 1 (blue), 10 (red). (E) Representative single-cell traces of relative fluorescence (F/F 0 ) of GCaMP5G (green) and mRuby (red) from cells infected by SA11-mRuby MOI 0.1 (purple) or 1 (blue). (F , G) Number of Ca 2+ spikes (F/F 0 > 5%) from mock or RV-infected cells that were infected with SA114F (F) or SA11-mRuby (G) . Data shown as mean ± SD of 60 cells/condition. **p < 0.01.

Article Snippet: Lentivirus constructs encoding shRNA targeting SA11 gene 10 and a non-targeting scrambled shRNA negative control were generated and packaged by America Pharma Source, LLC.

Techniques: Virus, Immunofluorescence, Infection, Staining, Recombinant, Fluorescence

Journal: Scientific Reports

Article Title: Efficient nucleic acid delivery to murine regulatory T cells by gold nanoparticle conjugates

doi: 10.1038/srep28709

Figure Lengend Snippet: Freshly isolated eGFP + regulatory T cells were cultured in presence or absence of 1.6 × 10 6 or 3.2 × 10 6 siRNA-coupled nanoparticle conjugates (5 nm) per cell or ligand-free particles as controls. eGFP expression was analyzed as mean fluorescence intensity (MFI) on gated living cells by flow cytometry ( A ) at day 2 or ( B ) day 3 and calculated as percentage of CD25 + untreated (= 100%). Results from three independent experiments are summarized as mean ± SEM. One-way ANOVA with Bonferroni´s post test was used for statistical analysis. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Successful transfection of T lymphocytes with plasmids or siRNA contructs was also achieved by others using transient plasmonic nanobubble injection or optimized electroporation protocols based on the Lonza nucleofection technique.

Techniques: Isolation, Cell Culture, Expressing, Fluorescence, Flow Cytometry

Journal: Cell Reports Medicine

Article Title: Myeloid cells coordinately induce glioma cell-intrinsic and cell-extrinsic pathways for chemoresistance via GP130 signaling

doi: 10.1016/j.xcrm.2024.101658

Figure Lengend Snippet: Humanin-induced chemoresistance requires ATR signaling (A) hGBM1 cells were stimulated with HN or vehicle (Ctrl.), underwent transcriptomics, and differentially expressed genes (DEGs) were analyzed by bioinformatics. (B) Experiments described in (A) were repeated with hGBM-1, 2, and 3 cells providing 12 consistent DEGs, of which several components assembled in a network. (C) HUS1 was associated with outcome in human GBMs. (D) In a myeloid-free brain sample, hGBMs have a basal level of HUS1 expression, which is upregulated by interaction with hiPSC microglia in a GP130-dependent manner. (E and F) Contribution of the ATR pathway to humanin-induced GBM expansion (E) and chemoresistance (F) was demonstrated with the ATR inhibitor AZ20. (G) Western blots showing expression levels of HUS1, ATR and beta-actin (loading control) and a readout for of ATR activation (pT1989) in hGBM1 cells treated with bovine serum albumin (control), TMZ, HN, or AZ20. (H) In summary, AZ20 does not cooperate with TMZ per se, but blocks HN-induced TMZ resistance. The number of biological replicates is indicated (dots in graphs indicate data from individual experiments); all error bars are presented as mean ± SDM. Statistical significance is shown as FDR in (A), one-way ANOVA (D, E), or two-way ANOVA (F): ∗ p < 0.05; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001; NS, not significant.

Article Snippet: HUS1 shRNA lentiviral and non-target control constructs , BioCat , Cat#: TLHSU1400-3364-pZIP-hCMV-ZsGreen-GVO-TRI.

Techniques: Expressing, Western Blot, Control, Activation Assay

Journal: Cell Reports Medicine

Article Title: Myeloid cells coordinately induce glioma cell-intrinsic and cell-extrinsic pathways for chemoresistance via GP130 signaling

doi: 10.1016/j.xcrm.2024.101658

Figure Lengend Snippet: Humanin-induced chemoresistance can be blocked therapeutically (A) Tumor size of orthotopic HN-WT or HN-C8a tumors was compared in mice receiving TMZ or vehicle (in animals with established tumor growth, 5x per week for 2 weeks; pre-defined endpoint was at 3 weeks). (B) Orthotopic hGBM1 was infused with HN (100 nM) or vehicle (artificial cerebrospinal fluid, aCSF) and i.p. injected with bazedoxifene-A (5 injections of BZA per week; 40 mg/kg; for 2 weeks) or vehicle; brains were labeled for HUS1; HUS1 expression was quantified. (C) Mice with established, orthotopic HN-WT tumors received TMZ (50 mg/kg) and were cotreated with vehicle or BZA (as in B); after 3 weeks, tumor size was quantified (dashed line: average data from untreated WT GBMs). (D) Mice with HN-WT GBMs received TMZ and were cotreated with vehicle or BZA (as in C); GBM samples were immunostained for active caspase-3 and immunolabeling was quantified (dashed line: average data from untreated WT GBMs). (E) Intratumoral vascularization and vessel diameter were compared in HN-WT or HN-C8a tumors receiving TMZ. (F) The HN-WT GBM mouse model was i.p. injected with TMZ and cotreated either with BZA or vehicle and the extent of intratumoral vascularization was compared. The number of biological replicates is indicated (dots in graphs indicate data from individual mice); all error bars are presented as mean ± SDM. Statistical significance is shown by one-way ANOVA (A, E), two-way ANOVA (B–D), or t test (F): ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; NS, not significant. Scale bars in (B, C) indicate 1 mm; scales in (D) represent 500 (overview) or 10 μm (magnified).

Article Snippet: HUS1 shRNA lentiviral and non-target control constructs , BioCat , Cat#: TLHSU1400-3364-pZIP-hCMV-ZsGreen-GVO-TRI.

Techniques: Injection, Labeling, Expressing, Immunolabeling

Journal: Cell Reports Medicine

Article Title: Myeloid cells coordinately induce glioma cell-intrinsic and cell-extrinsic pathways for chemoresistance via GP130 signaling

doi: 10.1016/j.xcrm.2024.101658

Figure Lengend Snippet:

Article Snippet: HUS1 shRNA lentiviral and non-target control constructs , BioCat , Cat#: TLHSU1400-3364-pZIP-hCMV-ZsGreen-GVO-TRI.

Techniques: Plasmid Preparation, Recombinant, Transfection, Fluorescence, Staining, Reverse Transcription, Expressing, Liposomes, Mutagenesis, shRNA, Control, Construct, Software, Imaging, Functional Assay, Dissection, Sequencing, Real-time Polymerase Chain Reaction

Journal: Oncotarget

Article Title: Hyperhomocysteinemia results from and promotes hepatocellular carcinoma via CYP450 metabolism by CYP2J2 DNA methylation

doi: 10.18632/oncotarget.14165

Figure Lengend Snippet: A, B . Representative chromatograms of 4 regioisomeric epoxyeicosatrienoic acids (EETs) in hepatocellular carcinoma (HCC) tumor and adjacent non-tumor tissue by LC/MS/MS assay. C . Left panel: Relative fold changes of 22 defined AA metabolites in 8 HCC tumor (T) and adjacent non-tumor (NT) tissue samples by LC/MS/MS assay. Right panel: Relative fold change of 11,12-EET levels in 42 pairs of HCC tumor (T) and adjacent non-tumor (NT) tissue by ELISA. D . Representative immunohistochemical staining of CYP2C8, CYP2C9, CYP2J2 and sEH in cross-sections of tumor and non-tumor tissue. Scale bars, 50 μm. E, F . qRT-PCR analysis of the relative mRNA expression of CYP2J2 and sEH in 42 cases of HCC tumor (T) and adjacent non-tumor (NT) tissue. β-actin expression was a normalization control. mRNA level of CYP2J2 and sEH in NT tissue was set to 1 for each paired HCC tumor tissue. G . The intercellular levels of Hcy in HCC tumor (T) and non-tumor (NT) tissue. H . 11,12-EET level in intracellular Hcy in HCC tissue. I . Correlation of Hcy concentration and CYP2J2 mRNA expression in HCC tissue. Data are mean±SD. *P<0.05 vs. NT.

Article Snippet: The siRNA targets CYP2J2, c-Jun and SP1, short hairpin RNA (sh-RNA) target CYP2J2, and corresponding controls were designed and synthesized by GenePharma (Shanghai).

Techniques: Liquid Chromatography with Mass Spectroscopy, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Staining, Quantitative RT-PCR, Expressing, Control, Concentration Assay

Journal: Oncotarget

Article Title: Hyperhomocysteinemia results from and promotes hepatocellular carcinoma via CYP450 metabolism by CYP2J2 DNA methylation

doi: 10.18632/oncotarget.14165

Figure Lengend Snippet: Intracellular levels of 11,12- and 14,15-EET A . and protein B . and mRNA C . levels of CYP2C8, CYP2C9, CYP2J2 and sEH by ELISA, western blot assay and quantitative RT-PCR, respectively, in LO2, SMMC7721 and HepG2 cells. Intercellular level of Hcy and folic acid (FA) D . and CYP2J2 mRNA E . in the above 3 cell lines, as well as 11,12-EET and 14,15-EET levels F . in SMMC7721 cells with Hcy and FA alone or combined. β-actin was an internal control. *P<0.05 vs. LO2 cells (A-C), * or # P<0.05 vs. corresponding control or Hcy treatment; $ P<0.05 vs. LO2 controls (D-F).

Article Snippet: The siRNA targets CYP2J2, c-Jun and SP1, short hairpin RNA (sh-RNA) target CYP2J2, and corresponding controls were designed and synthesized by GenePharma (Shanghai).

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Quantitative RT-PCR, Control

Journal: Oncotarget

Article Title: Hyperhomocysteinemia results from and promotes hepatocellular carcinoma via CYP450 metabolism by CYP2J2 DNA methylation

doi: 10.18632/oncotarget.14165

Figure Lengend Snippet: A . Structure of CpG sites (short vertical lines), CpG island (box), bisulfite sequencing region (dark bar) and transcription start sites (TSS) on the CYP2J2 promoter. Bisulfate genomic sequencing (BGS) analysis of CYP2J2 in SMMC7721, HepG2 and LO2 cells treated with or without 50 μM Hcy and 4 μM 5-aza for 72 hours. Each row indicates a clone from BGS to obtain a representative sampling of 32 CpG methylation patterns in the core promoter. Each circle corresponds to a single CpG site. Methylated sites are shown as filled circles, unmethylated sites as empty circles, and deletion or mutation sites as filled triangles. B, C . Bisulfite pyrosequencing analysis comparing the mean methylation for each CpG site (6th-14th) on the CYP2J2 promoter in HepG2 and LO2 cells treated with or without Hcy and in 42 HCC tumor (T) and adjacent non-tumor (NT) tissue samples. Data are mean±SD. *P<0.05 vs. LO2 cells (B) or 100% (C).

Article Snippet: The siRNA targets CYP2J2, c-Jun and SP1, short hairpin RNA (sh-RNA) target CYP2J2, and corresponding controls were designed and synthesized by GenePharma (Shanghai).

Techniques: Methylation Sequencing, Genomic Sequencing, Sampling, CpG Methylation Assay, Methylation, Mutagenesis

Journal: Oncotarget

Article Title: Hyperhomocysteinemia results from and promotes hepatocellular carcinoma via CYP450 metabolism by CYP2J2 DNA methylation

doi: 10.18632/oncotarget.14165

Figure Lengend Snippet: A . The SP1 and AP1 binding sites are underlined in the human CYP2J2 sequence. Serial 5’-deletion constructs of CYP2J2 promoter were co-transfected with CMV–β-gal into LO2, SMMC7721 and HepG2 cells for luciferase induction assay. *P<0.05 vs CYP2J2 -48/+100 bp-Luciferase reporter vector (Luc) activity in LO2 cells with PBS treatment; # P<0.05 vs luciferase activity of each construct in LO2 cells. B . AP1 or SP1 point-mutated constructs were generated on the basis of CYP2J2 -1000/+100 bp-Luc plasmids. Serial 5’-deletion constructs and mutated constructs were transiently co-transfected with CMV–β-gal (a transfection control) into LO2 cells and treated with Hcy or 5-aza for luciferase induction assay (relative to transfection with CYP2J2+30/−100 bp-Luc). *P<0.05 vs. CYP2J2 -48/+100 bp-Luc activity; # P<0.05 vs. Ctrl at each constructs. C . ChIP assay with anti-SP1 antibody for immunoprecipitation in LO2 (treated with Hcy or 5-aza, D.) SMMC7721 and HepG2 cells; normal rabbit IgG was a control. Semi-quantitative PCR with CYP2J2 promoter-specific primers to detect the binding of SP1 to the CYP2J2 promoter region. Shows quantification of the binding ability. Data are mean±SD (n=3). *P<0.05 vs. LO2 cells (C) or non-tumor tissue (D).

Article Snippet: The siRNA targets CYP2J2, c-Jun and SP1, short hairpin RNA (sh-RNA) target CYP2J2, and corresponding controls were designed and synthesized by GenePharma (Shanghai).

Techniques: Binding Assay, Sequencing, Construct, Transfection, Luciferase, Plasmid Preparation, Activity Assay, Generated, Control, Immunoprecipitation, Real-time Polymerase Chain Reaction

Journal: Oncotarget

Article Title: Hyperhomocysteinemia results from and promotes hepatocellular carcinoma via CYP450 metabolism by CYP2J2 DNA methylation

doi: 10.18632/oncotarget.14165

Figure Lengend Snippet: qRT-PCR and relative luciferase activity assay of SP1 A ., c-Jun B . and CYP2J2 mRNA expression and transcriptional activity (CYP2J2+1000/−100 bp-Luciferase reporter vector), respectively, with 40-nM siRNA targeting SP1 and c-Jun for 48 h: treatment with MAPK inhibitors (PD98059, U0126, SP600125 and SB203580) for 1 h with or without Hcy treatment in LO2 cells C, E . and HepG2 D, F . cells. β-actin was an internal control for qRT-PCR assay. Data are mean±SD (n=3). *P<0.05 vs. Ctrl; # P<0.05 vs. Hcy.

Article Snippet: The siRNA targets CYP2J2, c-Jun and SP1, short hairpin RNA (sh-RNA) target CYP2J2, and corresponding controls were designed and synthesized by GenePharma (Shanghai).

Techniques: Quantitative RT-PCR, Luciferase, Activity Assay, Expressing, Plasmid Preparation, Control

Journal: Oncotarget

Article Title: Hyperhomocysteinemia results from and promotes hepatocellular carcinoma via CYP450 metabolism by CYP2J2 DNA methylation

doi: 10.18632/oncotarget.14165

Figure Lengend Snippet: qRT-PCR A . and western blot B . assay of the mRNA and protein expression, respectively, of CYP2J2 in SMMC7721 cells with siRNA knockdown of CYP2J2 (Si-2J2) or control (Si-Ctrl) with or without Hcy treatment. C-E . The quantify of cell proliferation, clonogenicity, migration and invasion of SMMC7721 cells were measured by CCK-8 assay, flow cytometry, colony formation, tranwell assay, respectively. Data are mean±SD (n=3). *P<0.05 vs. Si-Ctrl; # P<0.05 vs. Hcy.

Article Snippet: The siRNA targets CYP2J2, c-Jun and SP1, short hairpin RNA (sh-RNA) target CYP2J2, and corresponding controls were designed and synthesized by GenePharma (Shanghai).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Knockdown, Control, Migration, CCK-8 Assay, Flow Cytometry

Journal: Oncotarget

Article Title: Hyperhomocysteinemia results from and promotes hepatocellular carcinoma via CYP450 metabolism by CYP2J2 DNA methylation

doi: 10.18632/oncotarget.14165

Figure Lengend Snippet: A . Targeting strategy and timeline of Hcy delivery in an orthotopic mouse model of HCC. B . Representative images of luciferase activity by the IVIS imaging system, tumor formation, and immunohistochemical staining for CYP2J2 at 8 weeks after luciferase-tagged HepG2 cells were injected into the left liver lobe of nude mice with different diets. C . Luminescence (cps) at 8, 10, 12, and 14 weeks. D-F . tumor weight (g) and CYP2J2 mRNA and protein expression. Methylation-specific PCR (MSP) analysis and ratio of DNA methylation to total methylation and unmethylation in liver of nude mice with different diets for 8 weeks. M: methylated, U: unmethylated. G . Western blot analysis of CYP2J2 protein expression with stable transfection with short hairpin CYP2J2 (sh-CYP2J2) and sh-Ctrl in HepG2 cells. Representative images of immunohistochemical staining (400x) of CYP2J2 H . and tumor weight I . at 8 weeks after HepG2 cells were transfected with sh-CYP2J2 and sh-Ctrl and injected into the left liver lobe of nude mice with Ctrl and M+ diet. Data are mean ± SD (n=8). *P<0.05 vs. Ctrl; # P<0.05 vs. M+. Scale bars, 50 μm.

Article Snippet: The siRNA targets CYP2J2, c-Jun and SP1, short hairpin RNA (sh-RNA) target CYP2J2, and corresponding controls were designed and synthesized by GenePharma (Shanghai).

Techniques: Luciferase, Activity Assay, Imaging, Immunohistochemical staining, Staining, Injection, Expressing, Methylation, DNA Methylation Assay, Western Blot, Stable Transfection, Transfection

Journal: Oncotarget

Article Title: Hyperhomocysteinemia results from and promotes hepatocellular carcinoma via CYP450 metabolism by CYP2J2 DNA methylation

doi: 10.18632/oncotarget.14165

Figure Lengend Snippet: Elevated Hcy level induces the accumulation of S-adenosyl homocysteine (SAH), which demethylates SP1 and AP1 (c-Jun/c-Fos) binding motifs on the CYP2J2 promoter via the ERK1/2 pathway. Combined effects increase CYP2J2 transcriptional activation and endogenous EET synthesis and promote primary tumor growth and metastasis.

Article Snippet: The siRNA targets CYP2J2, c-Jun and SP1, short hairpin RNA (sh-RNA) target CYP2J2, and corresponding controls were designed and synthesized by GenePharma (Shanghai).

Techniques: Binding Assay, Activation Assay