Journal: Neoplasia (New York, N.Y.)

Article Title: MiR-944/CISH mediated inflammation via STAT3 is involved in oral cancer malignance by cigarette smoking

doi: 10.1016/j.neo.2020.08.005

Figure Lengend Snippet: Phenotypic effects of dysregulated CISH in OSCC cell lines. (A) Western blot analysis of the STAT3, 5 and their phosphorylation form after transfection of control vector (Vec) or CISH expression vector (CISH) for 48 h in SAS and SCC-4 cells. GAPDH was used as protein loading control. Numerical values for protein band intensities are shown below the gels. The values were quantitated by densitometry and normalized to GAPDH. (B) The effect of CISH on the transcriptional activity of the construct containing the STAT3 binding sequence. The relative luciferase activities are the ratios of Renilla luciferase normalized to the vector alone control. (C) Migration assay following CISH overexpression for 24 h using Matrigel non-coated Boyden chamber assay in SAS and SCC-4 cells. (D) Invasion assay following CISH overexpression for 24 h using Matrigel coated Boyden chamber assay in SAS and SCC-4 cells. (E) CCL3, CCL5 and IL-1β levels in cultured medium of CISH overexpression SAS and SCC-4 cells measured by ELISA (compared with Vec control). (F) Western blot analysis of the CISH, COX-2 and iNOS after transfection of CISH for 48 h in SAS and SCC-4 cells. GAPDH was used as protein loading control. All data are represented as mean ± SD; **, p < 0.01; ***, p < 0.001.

Article Snippet: The membranes were probed with specific antibodies against CISH (#8731, Cell signaling, Danvers, MA), STAT3 (#610189, BD Biosciences, NJ), phospho-STAT3 (p-STAT3, Tyr 705 ) (#9131, Cell signaling), STAT5 (#25656, Cell signaling), phospho-STAT3 (p-STAT5, Tyr 694 ) (#9359, Cell signaling), COX-2 (#12282, Cell signaling), and iNOS (ab204017, Abcam, Cambridge, MA).

Techniques: Western Blot, Phospho-proteomics, Transfection, Control, Plasmid Preparation, Expressing, Activity Assay, Construct, Binding Assay, Sequencing, Luciferase, Migration, Over Expression, Boyden Chamber Assay, Invasion Assay, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Neoplasia (New York, N.Y.)

Article Title: MiR-944/CISH mediated inflammation via STAT3 is involved in oral cancer malignance by cigarette smoking

doi: 10.1016/j.neo.2020.08.005

Figure Lengend Snippet: CISH is a direct target of miR-944. (A) Flow chat of predicted miRNA which targets CISH 3′-UTR using two independent algorithms (microRNA.org and TargetScan) combined with our patients’ miRNA array data (GSE45238). (B) Western blot analysis of CISH protein after transfection of miR-944 mimics (PM) with 30 nM for 48 h in OEC-M1 and SCC-25 cells. (C) Schematic representation of the putative miR-944 binding sequence in the 3′-UTR of CISH with wild-type form (CISH 3′UTR WT) and mutant form (CISH 3′UTR Mut). The mutated nucleotides are labeled with underline. (D) Luciferase activity assays were performed after OEC-M1 and SCC-25 cells transfected with luciferase vectors containing the wide-type or mutant CISH 3′-UTR, in response to miR-944 over-expression. The relative luciferase activity of each sample is measured at 48 h after transfection and normalized to Renilla luciferase activity. (E) Correlation analysis of miR-944 and CISH in human OSCC patients ( n = 33) by qRT-PCR analysis. Western blot analysis of the CISH, STAT3 and phosphor-STAT3 after transfection of miR-944 mimics (PM) (F) or miR-944 inhibitors (AM) (G) with indicated concentration for 48 h in OEC-M1 and SCC-25 cells. All data are presented as mean ± SD; ***, p < 0.001 versus scramble control (NC). GAPDH was used as protein loading control. Numerical values for protein band intensities are shown below the gels. The values were quantitated by densitometry and normalized to GAPDH.

Article Snippet: The membranes were probed with specific antibodies against CISH (#8731, Cell signaling, Danvers, MA), STAT3 (#610189, BD Biosciences, NJ), phospho-STAT3 (p-STAT3, Tyr 705 ) (#9131, Cell signaling), STAT5 (#25656, Cell signaling), phospho-STAT3 (p-STAT5, Tyr 694 ) (#9359, Cell signaling), COX-2 (#12282, Cell signaling), and iNOS (ab204017, Abcam, Cambridge, MA).

Techniques: Western Blot, Transfection, Binding Assay, Sequencing, Mutagenesis, Labeling, Luciferase, Activity Assay, Over Expression, Quantitative RT-PCR, Concentration Assay, Control

Journal: Neoplasia (New York, N.Y.)

Article Title: MiR-944/CISH mediated inflammation via STAT3 is involved in oral cancer malignance by cigarette smoking

doi: 10.1016/j.neo.2020.08.005

Figure Lengend Snippet: miR-944 regulates OSCC cell functions through targeting CISH. OSCC cells were transfected with miR-944 mimics (PM) or scramble control (NC) for 24 h and then transfected with CISH expression vector without 3′-UTR (pCDNA3.1-CISH) or control vector (pCDNA3.1) for another 24 h. Under this condition, the level of CISH, STAT3 and phosphor-STAT3 were determined in OEC-M1 and SCC-25 cells by western blotting (A). The RNA level of COX-2 and iNOS (B), protein level of CCL3, CCL5 and IL-1β levels in cultured medium (C), migration ability (D), and invasion ability (E) were measured by qRT-PCR, ELISA and transwell assays in OEC-M1 cells. All data are presented as mean ± SD; **, p < 0.01; ***, p < 0.001 versus scramble control (NC). GAPDH was used as protein loading control. Numerical values for protein band intensities are shown below the gels. The values were quantitated by densitometry and normalized to GAPDH.

Article Snippet: The membranes were probed with specific antibodies against CISH (#8731, Cell signaling, Danvers, MA), STAT3 (#610189, BD Biosciences, NJ), phospho-STAT3 (p-STAT3, Tyr 705 ) (#9131, Cell signaling), STAT5 (#25656, Cell signaling), phospho-STAT3 (p-STAT5, Tyr 694 ) (#9359, Cell signaling), COX-2 (#12282, Cell signaling), and iNOS (ab204017, Abcam, Cambridge, MA).

Techniques: Transfection, Control, Expressing, Plasmid Preparation, Western Blot, Cell Culture, Migration, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Neoplasia (New York, N.Y.)

Article Title: MiR-944/CISH mediated inflammation via STAT3 is involved in oral cancer malignance by cigarette smoking

doi: 10.1016/j.neo.2020.08.005

Figure Lengend Snippet: NNK exposure induces miR-944 expression. (A) OEC-M1 and SCC-25 cells were treated with NNK (10–40 µM) for 72 h, and the miR-944 expression level was measured by qRT-PCR analysis. (B) OEC-M1 and SCC-25 cells were treated with 20 µM of NNK for indicated time (24–72 h), and the miR-944 expression level was measured by qRT-PCR analysis. (C) Luciferase activity assays were performed after OEC-M1 and SCC-25 cells transfected with luciferase vectors containing the wide-type or mutant CISH 3′-UTR, in response to DMSO control or 20 µM of NNK treatment. The relative luciferase activity of each sample is measured at 72 h after transfection and normalized to Renilla luciferase activity. OEC-M1 and SCC-25 cells were transfected with miR-944 inhibitors (AM) for 12 h, and followed by 20 µM of NNK treatment for another 72 h. Under this condition, the expression level of miR-944 was determined by qRT-PCR analysis (D), and the protein level of CISH, STAT3, phosphor-STAT3 and GAPDH were determined by western blot analysis (E). All data are presented as mean ± SD; *, p < 0.05; **, p < 0.01; ***, p < 0.001. Numerical values for protein band intensities are shown below the gels. The values were quantitated by densitometry and normalized to GAPDH.

Article Snippet: The membranes were probed with specific antibodies against CISH (#8731, Cell signaling, Danvers, MA), STAT3 (#610189, BD Biosciences, NJ), phospho-STAT3 (p-STAT3, Tyr 705 ) (#9131, Cell signaling), STAT5 (#25656, Cell signaling), phospho-STAT3 (p-STAT5, Tyr 694 ) (#9359, Cell signaling), COX-2 (#12282, Cell signaling), and iNOS (ab204017, Abcam, Cambridge, MA).

Techniques: Expressing, Quantitative RT-PCR, Luciferase, Activity Assay, Transfection, Mutagenesis, Control, Western Blot

Journal: Neoplasia (New York, N.Y.)

Article Title: MiR-944/CISH mediated inflammation via STAT3 is involved in oral cancer malignance by cigarette smoking

doi: 10.1016/j.neo.2020.08.005

Figure Lengend Snippet: The effects of NNK on the STAT3-mediated pro-inflammation genes. OEC-M1 and SCC-25 cells were transfected with miR-944 inhibitors (AM) for 12 h, and followed by 20 µM of NNK treatment for another 72 h. The RNA level of COX-2 and iNOS in cells (A), protein level of CCL3, CCL5 and IL-1β in cultured medium (B) were measured by qRT-PCR or ELISA. (C) Migration and invasion ability were measured by transwell assays in OEC-M1 and SCC-25 cells. (D) OEC-M1 and SCC-25 cells were treated with 20 µM of NNK for 72 h, and the TP63 expression level was measured by qRT-PCR analysis. (E) Correlation analysis of miR-944 and TP63 in human OSCC patients (n = 33) by qRT-PCR analysis. (F) Proposed model for comprehensive NNK-induced upregulation of miR-944 in stimulating STAT3 activation and pro-inflammatory genes expression through suppression of CISH. All data are represented as mean ± SD; **, p < 0.01; ***, p < 0.001.

Article Snippet: The membranes were probed with specific antibodies against CISH (#8731, Cell signaling, Danvers, MA), STAT3 (#610189, BD Biosciences, NJ), phospho-STAT3 (p-STAT3, Tyr 705 ) (#9131, Cell signaling), STAT5 (#25656, Cell signaling), phospho-STAT3 (p-STAT5, Tyr 694 ) (#9359, Cell signaling), COX-2 (#12282, Cell signaling), and iNOS (ab204017, Abcam, Cambridge, MA).

Techniques: Transfection, Cell Culture, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Migration, Expressing, Activation Assay

Journal: Pharmacological research

Article Title: Ectopic expression of NKG7 enhances CAR-T function and improves the therapeutic efficacy in liquid and solid tumors.

doi: 10.1016/j.phrs.2024.107506

Figure Lengend Snippet: Fig. 4. NKG7 promotes the proliferation of CAR-T cells by enhancing the IL-2 signaling pathway. (A and B) GSEA of DNA replication and cell cycle between B7H3- CAR-TNKG7 and B7H3-CAR-T cells following B7H3 protein stimulation for 6 h. (C) Volcano plot of T cell proliferation-related genes between B7H3-CAR-TNKG7 and B7H3-CAR-T cells. (D) Heatmaps of significant T cell proliferation-related genes. (E and F) IL-2 expression of CAR-T cells was measured by FCM analysis following target cell stimulation for 6 h. (G) Statistics for (E and F). (H) The proliferation of CAR-T cells was detected by a CFSE assay. (I and J) GSEA of the Jak-STAT and PI3K- Akt pathways between B7H3-CAR-TNKG7 and B7H3-CAR-T cells following B7H3 protein stimulation for 6 h. (K) The IL-2 signaling pathway (p-AKT (Ser473), AKT, p- ERK (Thr202/Tyr204), ERK, p-STAT5 (Thr694) and STAT5) in CAR-T cells were explored by WB assays. ***P < 0.001.

Article Snippet: After blocking with 5 % nonfat milk at room temperature for 1 h, the membranes were incubated with primary antibodies against NKG7 (Cat No. 84835, 1:1000, CST), AKT (Cat No. A11016, 1:1000, ABclonal), p-AKT (Ser473) (Cat No. AP0140, 1:1000, ABclonal), ERK (Cat No. 4695, 1:1000, CST), p-ERK (Thr202/Tyr204) (Cat No. 4370, 1:1000, CST), STAT5 (Cat No. 25656, 1:1000, CST), pSTAT5 (Thr694) (Cat No. 4322, 1:1000, CST) and β-actin (Cat No. 66009–1-Ig, 1:5000, Proteintech) overnight at 4◦C.

Techniques: Expressing, Cell Stimulation, CFSE Assay

Journal: International Journal of Molecular Sciences

Article Title: Different Lipid Regulation in Ovarian Cancer: Inhibition of the Immune System

doi: 10.3390/ijms19010273

Figure Lengend Snippet: Normal expression and phosphorylation of the IL-2 receptor and its downstream signaling components. PBLs of healthy individuals or sorted ascites-derived CD4 + CD25 − T cells (5 × 10 5 cells/well) were stimulated with 20,000 αCD3/28 beads/well for three days. Subsequently, samples were stained for flow cytometry analysis. ( A ) Cells were stained for isotype control, CD25, CD122, and CD132. Plots are representative for eight controls and three patients. ( B ) Control ( n = 3) and ascites-derived T cells ( n = 3) were stained for JAK3, STAT5, phospho-STAT5, phosphor-AKT and phosphor-ERK. One representative example is shown.

Article Snippet: The following antibodies were used in this study: CD4-APC (300514, Beckman Coulter, Brea, CA, USA), CD4-FITC (A07750, Beckman Coulter), CD25-PE (555432, BD Biosciences, San Jose, CA, USA), CD122-PE (IM1978, Beckman Coulter), CD132-APC (338608, BioLegend, San Diego, CA, USA), cyclin D (2936P, Cell Signaling, Danvers, MA, USA), cyclin E (551159, Cell Signaling, Danvers, MA, USA), JAK3 (ab45141, Abcam, Cambridge, UK), STAT5 (9363P, Cell Signaling, Danvers, MA, USA), pSTAT5 (9314S, Cell Signaling, Danvers, MA, USA), pAKT (4075S, Cell Signaling, Danvers, MA, USA), and pERK1/2 (4284S, Cell Signaling).

Techniques: Expressing, Phospho-proteomics, Derivative Assay, Staining, Flow Cytometry, Control

Journal: Redox Biology

Article Title: Bi-directionally protective communication between neurons and astrocytes under ischemia

doi: 10.1016/j.redox.2017.05.010

Figure Lengend Snippet: Involvement of ERK, PI3K and STAT5 in EPO-mediated anti-apoptosis in astrocytes. In A (n = 10) and B (n = 6), astrocytes were incubated with IPc-0 h NCM (IPc-0h) with or without ischemia (Isch), or with IPc-1h in the presence of AG490 (AG), PD98059 (PD), LY294001 (LY), 573108 (STAT5 inhibitor), BAY11-7082 (BAY) and SB203850 (SB) for 1-h, followed with IPc-1h NCM for 48-h and exposure to ischemia for 12-h. Viabilities were then assayed. In C to F , astrocytes were incubated in IPc-1h NCM (IPc-1h) with or without anti-EPO antibody (aEPO) for 20 min, and p-Akt ( C ), p-Erk ( D ), p-STAT3 ( E ) and p-STAT5 ( F ) were then measured by Western blot (n = 3). G–J : Astrocytes were incubated in IPc-1h NCM with or without aEPO for 48-h, and Bcl-2 ( G ), 136 p-Bad ( H ), 112 p-Bad ( I ) and Bcl-xL ( J ) were then measured by Western blot (n = 3). K-N : Astrocytes were pre-treated with or without PD98059, LY294002 or 573108 for 1-h before being incubated in IPc-1h NCM for 48-h, and Bcl-2 ( K ), 136 p-Bad ( L ), 112 p-Bad ( M ) and Bcl-xL ( N ) were then measured by Western blot (n = 3). Data were Mean ±SD. *p < 0.05, **p < 0.01,***p < 0.001. versus the corresponding control (Isch = Ischemia).

Article Snippet: Primary polyclonal rabbit anti-Akt, phosphorylated Akt ( p -Akt), extracellular signal-regulated kinase (ERK) 42/44, phosphorylated ERK 42/44 (p-ERK42/44), Bad, phosphorylated Bad Ser 112 (112p-Bad), phosphorylated Bad Ser 136 (136p-Bad), Bcl-2, cleaved caspase-3, caspase-3, phosphorylated signal transducer and activator of transcription 3 (p-STAT3) and 5 (p-STAT5) antibodies were purchased from Cell Signaling Technology, Beverly, MA, USA; primary monoclonal mouse anti-hypoxia-inducible factor-1 alpha (HIF-1 alpha) antibody from Novus Biologicals, Inc., Littleton, CO, USA; primary polyclonal rabbit anti-EPO antibody from Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA; antibodies against neuron microtubule-associated protein 2 (MAP2) and astrocyte glial fibrillary acadic protein (GFAP) from Chemicon International Ltd, Hampshire, UK; and both mouse anti-Histone 3 monoclonal antibody and Ab175819-8 isoprostane from Abcam, Cambridge, UK.

Techniques: Incubation, Western Blot, Control

Journal: Redox Biology

Article Title: Bi-directionally protective communication between neurons and astrocytes under ischemia

doi: 10.1016/j.redox.2017.05.010

Figure Lengend Snippet: A hypothetical scheme for the mechanisms involved in the astro-protection by ischemia-preconditioned neurons. IP neurons have the ability to express and release sufficient amounts of EPO for paracrine astro-protection via the increased HIF-1 alpha induced by IP. The increased EPO binds to EPOR on the membrane of astrocytes, triggering dimerization of EPOR and inducing Jak2 activation, which in turn activates three different downstream-signaling pathways to prevent apoptosis: PI3K/Akt, ERK and STAT5. Activated PI3K/Akt can phosphorylate Bad at ser-136 and up-regulate Bcl-2 expression, and in turn preventing apoptosis. ERK signaling pathway-mediated anti-apoptosis is via enhanced Bcl-2 levels and phosphorylation of Bad at ser-112, while STAT5 suppresses astrocyte apotosis via the up-regulated expression of Bcl-xL. The anti-apoptotic effects of EPO from IP neurons on astrocytes require the combined activation of these three pathways. In addition, IP neurons were able to inhibit I/R induced-oxidative stress by up-regulation of anti-oxidant enzymes (SOD, CAT, GSH-PX) partly via EPO. Therefore. IP neurons protect astrocytes from ischemia-induced injury by an EPO-mediated anti-apoptosis and anti-oxidant effect.

Article Snippet: Primary polyclonal rabbit anti-Akt, phosphorylated Akt ( p -Akt), extracellular signal-regulated kinase (ERK) 42/44, phosphorylated ERK 42/44 (p-ERK42/44), Bad, phosphorylated Bad Ser 112 (112p-Bad), phosphorylated Bad Ser 136 (136p-Bad), Bcl-2, cleaved caspase-3, caspase-3, phosphorylated signal transducer and activator of transcription 3 (p-STAT3) and 5 (p-STAT5) antibodies were purchased from Cell Signaling Technology, Beverly, MA, USA; primary monoclonal mouse anti-hypoxia-inducible factor-1 alpha (HIF-1 alpha) antibody from Novus Biologicals, Inc., Littleton, CO, USA; primary polyclonal rabbit anti-EPO antibody from Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA; antibodies against neuron microtubule-associated protein 2 (MAP2) and astrocyte glial fibrillary acadic protein (GFAP) from Chemicon International Ltd, Hampshire, UK; and both mouse anti-Histone 3 monoclonal antibody and Ab175819-8 isoprostane from Abcam, Cambridge, UK.

Techniques: Membrane, Activation Assay, Protein-Protein interactions, Expressing, Phospho-proteomics

Journal: Nature Communications

Article Title: NKp46 enhances type 1 innate lymphoid cell proliferation and function and anti-acute myeloid leukemia activity

doi: 10.1038/s41467-025-55923-w

Figure Lengend Snippet: a Ncr1 +/+ and Ncr1 gfp/gfp ILC1s were sorted for RNA-seq from the liver of Ncr1 +/+ mice and Ncr1 gfp/gfp mice, respectively, using FACS. Volcano plots show statistically significant differentially expressed genes in RNA pools ( Ncr1 gfp/gfp ILC1s vs. Ncr1 +/+ ILC1s) ( n = 5). b – d Expression of IL-18R1 ( b ), IL-7Rα ( c ), and IL-2Rα ( d ) on the liver Ncr1 +/+ ILC1s and Ncr1 gfp/gfp ILC1s was measured by flow cytometry ( n = 6). e Ncr1 +/+ ILC1s and Ncr1 gfp/gfp ILC1s were treated with IL-18 (10 ng/ml) at the indicated time points. Phosphorylation of P38 was measured by flow cytometry ( Ncr1 +/+ : n = 5; Ncr1 gfp/gfp : n = 4). f , g Ncr1 +/+ ILC1s and Ncr1 gfp/gfp ILC1s were treated with IL-7 (100 ng/ml; f ) or IL-2 (1000 IU/ml; g ) at the indicated time points. Phosphorylation of STAT5 was measured by flow cytometry ( f : n = 7; g : n = 5). Data were presented as means ± s.d.; P values were calculated by either a two-tailed Student’s t -test ( b – d and f at 30 min) or linear mixed models with adjustments ( e , g ). Source data are provided as a Source Data file.

Article Snippet: They were then stained with p-STAT3 (BV421-conjugated anti-p-STAT3, BioLegend, Cat# 651010, Clone: 13A3-1), p-STAT5 (PE-conjugated anti-p-STAT5, Cell Signaling Technology, Cat# 14603, Clone: D47E7), or p-P65 (PE-conjugated anti-p-P65, Cell Signaling Technology, Cat# 653004, Clone: 14G10A21) antibodies in flow cytometry buffer.

Techniques: RNA Sequencing, Expressing, Flow Cytometry, Phospho-proteomics, Two Tailed Test

Journal: eLife

Article Title: Unsupervised machine learning reveals risk stratifying glioblastoma tumor cells

doi: 10.7554/eLife.56879

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-p-STAT5 (Y694) (mouse-monoclonal) , Fluidigm , RRID: AB_2744690 Cat#3150005A Clone:47 , MC (1:100).

Techniques: Isolation, Software

Journal: eLife

Article Title: HDAC3 restrains CD8-lineage genes to maintain a bi-potential state in CD4 + CD8 + thymocytes for CD4-lineage commitment

doi: 10.7554/eLife.43821

Figure Lengend Snippet: ( A ) Surface expression of cytokine receptor chains γc, IL-4Rα, IL-7Rα, IL-15Rα, IL-2Rβ, and IL-21R on DP thymocytes from WT and HDAC3-cKO mice. Flow cytometric plots have an isotype control to illustrate background level of expression. ( B ) Gene expression (RNA-seq) of Il2rg , Il4ra, Il7r, Il15ra, Il2rb , and Il21r in Immature (DP) cells from OT-II and OT-II HDAC3-cKO mice. ( C ) Snapshot of H3K27ac ChIP-seq tracks at the Il4ra and Il21r gene loci in Immature (DP) thymocytes from OT-II and OT-II HDAC3-cKO mice. Shaded regions identify super-enhancers. ( D ) Protein expression of IL-21R on DP, CD4SP and CD8SP thymocytes from WT mice and DP thymocytes from HDAC3-cKO mice. ( E ) pSTAT5 levels in DP thymocytes from WT and HDAC3-cKO mice after 10 min ex vivo stimulation with IL-21, IL-15, IL-7, IL4, or media alone. ( F ) Protein expression of STAT5a and STAT5b in DP thymocytes from WT and HDAC3-cKO mice. ( G ) pSTAT3 and pSTAT1 levels after a 10 min in vitro IL-21 stimulation, and pSTAT6 levels after IL-4 stimulation of DP thymocytes from WT and HDAC3-cKO mice. ( H ) Quantitative ChIP (qChIP) of HDAC3 binding at the Il21r promoter in DP thymocytes from WT and HDAC3-cKO mice. Graph depicts fold enrichment over Rpl30 (n = 4 mice/group from four indpendent experiments). ( I ) DNase-seq and Hi-C arc plots at the Il4ra and Il21r gene loci in DP thymocytes and pooled DN3-to-DP thymocytes, respectively. Shaded region highlights where HDAC3 binds, as shown in . (DHS, DNA hypersensitivity sites). ( A, E, G ) Bar graph shows mean ± SEM of MFI from 4 to 5 mice from at least three independent experiments. ( D, H ) Plots are representative of at least three mice from three independent experiments. ( F ) Bar graph shows mean ± SEM of MFI from three mice from two independent experiments. (***, p < 0.001). See also – .

Article Snippet: After stimulation, cells were immediately fixed with BD Lyse/Fix Buffer (BD Phosflow kit), stained for surface proteins, permeabolized with BD Perm Buffer III (BD Phosflow kit), and stained with antibodies targeting pSTAT5 (#9365, Cell Signaling Technology), pSTAT1 (#8009, Cell Signaling Technology), pSTAT3 (#4324, Cell Signaling Technology), or pSTAT6 (#9361, Cell Signaling Technology) for 30 min at room temperature.

Techniques: Expressing, Control, Gene Expression, RNA Sequencing, ChIP-sequencing, Ex Vivo, In Vitro, Binding Assay, Hi-C