m1 protein Search Results


sino biological cat 40010 v07e  (Sino Biological)
94
Sino Biological sino biological cat 40010 v07e
Sino Biological Cat 40010 V07e, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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foxm1  (MedChemExpress)
93
MedChemExpress foxm1
Foxm1, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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pkm2  (Proteintech)
96
Proteintech pkm2
Fig. 7. Astragalin (AST) eliminated lipopolysaccharide (LPS)-induced inflammatory response by inhibiting glycolysis enhancement mediated by the hypoxia- inducible factor-1α/pyruvate kinase M2 <t>(HIF-1α/PKM2)</t> signaling pathway. (A) LPS changed the concentrations of adenosine triphosphate (ATP) and lactic acid (LA) in liver and serum, and AST reversed these changes. Student’s t-test was performed, n = 4, * P < 0.05, ** P < 0.01, *** P < 0.001. (B) The activities of hexokinase (HK), pyruvate kinase (PK), and lactate dehydrogenase (LDH) in liver and serum were enhanced by LPS, whereas AST inhibited the enhancement. Student’s t-test was performed, n = 4, * P < 0.05, ** P < 0.01, *** P < 0.001. (C) Glycolysis-related genes, including HK2, LDHA, glucose-6-phosphate dehydrogenase X-linked (G6pdx), and glucose transporter 1 (Glut-1), were upregulated by LPS but AST prevented the upregulation. Similar results also showed in the expression of HIF-1α and PKM2 genes. Student’s t-test was performed, n = 4, * P < 0.05, ** P < 0.01, *** P < 0.001. (D-E) Expressions of HIF-1α and PKM2 proteins were increased by LPS treatment, while AST decreased their expression. Student’s t-test was performed, n = 3, * P < 0.05, ** P < 0.01.
Pkm2, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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pkm 201  (Proteintech)
93
Proteintech pkm 201
Fig. 7. Astragalin (AST) eliminated lipopolysaccharide (LPS)-induced inflammatory response by inhibiting glycolysis enhancement mediated by the hypoxia- inducible factor-1α/pyruvate kinase M2 <t>(HIF-1α/PKM2)</t> signaling pathway. (A) LPS changed the concentrations of adenosine triphosphate (ATP) and lactic acid (LA) in liver and serum, and AST reversed these changes. Student’s t-test was performed, n = 4, * P < 0.05, ** P < 0.01, *** P < 0.001. (B) The activities of hexokinase (HK), pyruvate kinase (PK), and lactate dehydrogenase (LDH) in liver and serum were enhanced by LPS, whereas AST inhibited the enhancement. Student’s t-test was performed, n = 4, * P < 0.05, ** P < 0.01, *** P < 0.001. (C) Glycolysis-related genes, including HK2, LDHA, glucose-6-phosphate dehydrogenase X-linked (G6pdx), and glucose transporter 1 (Glut-1), were upregulated by LPS but AST prevented the upregulation. Similar results also showed in the expression of HIF-1α and PKM2 genes. Student’s t-test was performed, n = 4, * P < 0.05, ** P < 0.01, *** P < 0.001. (D-E) Expressions of HIF-1α and PKM2 proteins were increased by LPS treatment, while AST decreased their expression. Student’s t-test was performed, n = 3, * P < 0.05, ** P < 0.01.
Pkm 201, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse monoclonal anti m1  (Bio-Rad)
93
Bio-Rad mouse monoclonal anti m1
(A) Diagram of quadruple baculovirus transfer vector encoding influenza virus genes HA, NA, <t>M1,</t> and M2. This vector was transfected together with linearized baculovirus DNA into Sf9 insect cells to generate the quadruple recombinant (HA/Q28). Replacement of the HA with a gene encoding a full-length sequence of the VSV-G protein or chimera VSV-G/HA (see below) generated VSV-G/Q or VSV-G/HA/Q, respectively. (B) Structure of the VSV-G/HA chimera. The 11 aa of the cytoplasmic tail, 26 aa of the transmembrane domain, and 5 aa of the ectodomain of the influenza virus HA were fused in frame with the ectodomain of the VSV-G surface glycoprotein. This hybrid gene was used to replace the HA from the quadruple transfer vector (panel A) and generate the VSV-G/HA/Q gene.
Mouse Monoclonal Anti M1, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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m1 protein  (Sino Biological)
94
Sino Biological m1 protein
Deletion <t>of</t> <t>Suv4-20h2</t> enhances influenza viral replication (A) WT mice were intratracheally infected with influenza virus (PR8 virus, 100 FFU), and lung tissues were sampled at the indicated time points. Histone was extracted from the lung tissues and was subjected to H4-tail proteomics analysis. Relative expression levels of H4K20me1, H4K20me2, and H4K20me3 to H4 were shown (n = 5 per group). (B–D) MEFs obtained from WT, Suv4-20h1 KO (h1 KO), Suv4-20h2 KO (h2 KO) and Suv4-20h1/Suv4-20h2 double KO (dKO) mice were infected with mock (Flu-) or influenza virus (MOI: 0.5) (Flu+). Cells were fixed and stained with either H4K20me1, H4K20me2 or H4K20me3 Abs, and Hoechst. Data are from three separate experiments. Representative staining is shown. Scale bars: 10 μm (B). Culture supernatant and cells were sampled at 8 hr after infection. Virus titer assessed by FFU in supernatant is shown (C, upper panel). NP mRNA expression of the cells is shown (C, lower panel). Data are from three separate experiments. ∗p < 0.05 compared to WT. Infected cells were fixed, and virus NP protein was stained (green). Nuclei were counterstained with Hoechst (blue). Representative staining is shown. Scale bars: 50 μm (D, left panel). The percentages of NP-positive in total of Hoechst positive are shown (D, right panel). ∗∗p < 0.01 compared to WT. (E) WT and dKO MEFs were infected with influenza virus (MOI: 0.5) and sampled at the indicated time point. Western blots of virus PB2, NP and <t>M1</t> proteins, along with GAPDH are shown. Data were reproduced in three experiments. The data in (A, C and D) (right panel) are presented as means ± s.e.m.. Statistical analysis were carried out using analysis of variance with Bonferroni post t tests.
M1 Protein, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit polyclonal anti m1 antibody  (Sino Biological)
94
Sino Biological rabbit polyclonal anti m1 antibody
Deletion <t>of</t> <t>Suv4-20h2</t> enhances influenza viral replication (A) WT mice were intratracheally infected with influenza virus (PR8 virus, 100 FFU), and lung tissues were sampled at the indicated time points. Histone was extracted from the lung tissues and was subjected to H4-tail proteomics analysis. Relative expression levels of H4K20me1, H4K20me2, and H4K20me3 to H4 were shown (n = 5 per group). (B–D) MEFs obtained from WT, Suv4-20h1 KO (h1 KO), Suv4-20h2 KO (h2 KO) and Suv4-20h1/Suv4-20h2 double KO (dKO) mice were infected with mock (Flu-) or influenza virus (MOI: 0.5) (Flu+). Cells were fixed and stained with either H4K20me1, H4K20me2 or H4K20me3 Abs, and Hoechst. Data are from three separate experiments. Representative staining is shown. Scale bars: 10 μm (B). Culture supernatant and cells were sampled at 8 hr after infection. Virus titer assessed by FFU in supernatant is shown (C, upper panel). NP mRNA expression of the cells is shown (C, lower panel). Data are from three separate experiments. ∗p < 0.05 compared to WT. Infected cells were fixed, and virus NP protein was stained (green). Nuclei were counterstained with Hoechst (blue). Representative staining is shown. Scale bars: 50 μm (D, left panel). The percentages of NP-positive in total of Hoechst positive are shown (D, right panel). ∗∗p < 0.01 compared to WT. (E) WT and dKO MEFs were infected with influenza virus (MOI: 0.5) and sampled at the indicated time point. Western blots of virus PB2, NP and <t>M1</t> proteins, along with GAPDH are shown. Data were reproduced in three experiments. The data in (A, C and D) (right panel) are presented as means ± s.e.m.. Statistical analysis were carried out using analysis of variance with Bonferroni post t tests.
Rabbit Polyclonal Anti M1 Antibody, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit anti m1  (Sino Biological)
92
Sino Biological rabbit anti m1
Deletion <t>of</t> <t>Suv4-20h2</t> enhances influenza viral replication (A) WT mice were intratracheally infected with influenza virus (PR8 virus, 100 FFU), and lung tissues were sampled at the indicated time points. Histone was extracted from the lung tissues and was subjected to H4-tail proteomics analysis. Relative expression levels of H4K20me1, H4K20me2, and H4K20me3 to H4 were shown (n = 5 per group). (B–D) MEFs obtained from WT, Suv4-20h1 KO (h1 KO), Suv4-20h2 KO (h2 KO) and Suv4-20h1/Suv4-20h2 double KO (dKO) mice were infected with mock (Flu-) or influenza virus (MOI: 0.5) (Flu+). Cells were fixed and stained with either H4K20me1, H4K20me2 or H4K20me3 Abs, and Hoechst. Data are from three separate experiments. Representative staining is shown. Scale bars: 10 μm (B). Culture supernatant and cells were sampled at 8 hr after infection. Virus titer assessed by FFU in supernatant is shown (C, upper panel). NP mRNA expression of the cells is shown (C, lower panel). Data are from three separate experiments. ∗p < 0.05 compared to WT. Infected cells were fixed, and virus NP protein was stained (green). Nuclei were counterstained with Hoechst (blue). Representative staining is shown. Scale bars: 50 μm (D, left panel). The percentages of NP-positive in total of Hoechst positive are shown (D, right panel). ∗∗p < 0.01 compared to WT. (E) WT and dKO MEFs were infected with influenza virus (MOI: 0.5) and sampled at the indicated time point. Western blots of virus PB2, NP and <t>M1</t> proteins, along with GAPDH are shown. Data were reproduced in three experiments. The data in (A, C and D) (right panel) are presented as means ± s.e.m.. Statistical analysis were carried out using analysis of variance with Bonferroni post t tests.
Rabbit Anti M1, supplied by Sino Biological, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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influenza a  (Sino Biological)
91
Sino Biological influenza a
Deletion <t>of</t> <t>Suv4-20h2</t> enhances influenza viral replication (A) WT mice were intratracheally infected with influenza virus (PR8 virus, 100 FFU), and lung tissues were sampled at the indicated time points. Histone was extracted from the lung tissues and was subjected to H4-tail proteomics analysis. Relative expression levels of H4K20me1, H4K20me2, and H4K20me3 to H4 were shown (n = 5 per group). (B–D) MEFs obtained from WT, Suv4-20h1 KO (h1 KO), Suv4-20h2 KO (h2 KO) and Suv4-20h1/Suv4-20h2 double KO (dKO) mice were infected with mock (Flu-) or influenza virus (MOI: 0.5) (Flu+). Cells were fixed and stained with either H4K20me1, H4K20me2 or H4K20me3 Abs, and Hoechst. Data are from three separate experiments. Representative staining is shown. Scale bars: 10 μm (B). Culture supernatant and cells were sampled at 8 hr after infection. Virus titer assessed by FFU in supernatant is shown (C, upper panel). NP mRNA expression of the cells is shown (C, lower panel). Data are from three separate experiments. ∗p < 0.05 compared to WT. Infected cells were fixed, and virus NP protein was stained (green). Nuclei were counterstained with Hoechst (blue). Representative staining is shown. Scale bars: 50 μm (D, left panel). The percentages of NP-positive in total of Hoechst positive are shown (D, right panel). ∗∗p < 0.01 compared to WT. (E) WT and dKO MEFs were infected with influenza virus (MOI: 0.5) and sampled at the indicated time point. Western blots of virus PB2, NP and <t>M1</t> proteins, along with GAPDH are shown. Data were reproduced in three experiments. The data in (A, C and D) (right panel) are presented as means ± s.e.m.. Statistical analysis were carried out using analysis of variance with Bonferroni post t tests.
Influenza A, supplied by Sino Biological, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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gene exp lamp1 mm00495262 m1  (Thermo Fisher)
94
Thermo Fisher gene exp lamp1 mm00495262 m1
Deletion <t>of</t> <t>Suv4-20h2</t> enhances influenza viral replication (A) WT mice were intratracheally infected with influenza virus (PR8 virus, 100 FFU), and lung tissues were sampled at the indicated time points. Histone was extracted from the lung tissues and was subjected to H4-tail proteomics analysis. Relative expression levels of H4K20me1, H4K20me2, and H4K20me3 to H4 were shown (n = 5 per group). (B–D) MEFs obtained from WT, Suv4-20h1 KO (h1 KO), Suv4-20h2 KO (h2 KO) and Suv4-20h1/Suv4-20h2 double KO (dKO) mice were infected with mock (Flu-) or influenza virus (MOI: 0.5) (Flu+). Cells were fixed and stained with either H4K20me1, H4K20me2 or H4K20me3 Abs, and Hoechst. Data are from three separate experiments. Representative staining is shown. Scale bars: 10 μm (B). Culture supernatant and cells were sampled at 8 hr after infection. Virus titer assessed by FFU in supernatant is shown (C, upper panel). NP mRNA expression of the cells is shown (C, lower panel). Data are from three separate experiments. ∗p < 0.05 compared to WT. Infected cells were fixed, and virus NP protein was stained (green). Nuclei were counterstained with Hoechst (blue). Representative staining is shown. Scale bars: 50 μm (D, left panel). The percentages of NP-positive in total of Hoechst positive are shown (D, right panel). ∗∗p < 0.01 compared to WT. (E) WT and dKO MEFs were infected with influenza virus (MOI: 0.5) and sampled at the indicated time point. Western blots of virus PB2, NP and <t>M1</t> proteins, along with GAPDH are shown. Data were reproduced in three experiments. The data in (A, C and D) (right panel) are presented as means ± s.e.m.. Statistical analysis were carried out using analysis of variance with Bonferroni post t tests.
Gene Exp Lamp1 Mm00495262 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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tenm1  (Proteintech)
92
Proteintech tenm1
Identification of the ECM-related genes associated with patients survival. Notes . (a) Univariate Cox regression analysis was utilized to identify the prognosis-related genes with P < 0.05; (b) LASSO regression analysis; (c) multivariate cox regression analysis based on the genes screened by LASSO regression; (d–i) clinical correlation of IBSP, LINGO4, COL26A1, MMP12, KLK4, RTBDN, <t>TENM1,</t> GDF15, and RUNX1.
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Image Search Results


Fig. 7. Astragalin (AST) eliminated lipopolysaccharide (LPS)-induced inflammatory response by inhibiting glycolysis enhancement mediated by the hypoxia- inducible factor-1α/pyruvate kinase M2 (HIF-1α/PKM2) signaling pathway. (A) LPS changed the concentrations of adenosine triphosphate (ATP) and lactic acid (LA) in liver and serum, and AST reversed these changes. Student’s t-test was performed, n = 4, * P < 0.05, ** P < 0.01, *** P < 0.001. (B) The activities of hexokinase (HK), pyruvate kinase (PK), and lactate dehydrogenase (LDH) in liver and serum were enhanced by LPS, whereas AST inhibited the enhancement. Student’s t-test was performed, n = 4, * P < 0.05, ** P < 0.01, *** P < 0.001. (C) Glycolysis-related genes, including HK2, LDHA, glucose-6-phosphate dehydrogenase X-linked (G6pdx), and glucose transporter 1 (Glut-1), were upregulated by LPS but AST prevented the upregulation. Similar results also showed in the expression of HIF-1α and PKM2 genes. Student’s t-test was performed, n = 4, * P < 0.05, ** P < 0.01, *** P < 0.001. (D-E) Expressions of HIF-1α and PKM2 proteins were increased by LPS treatment, while AST decreased their expression. Student’s t-test was performed, n = 3, * P < 0.05, ** P < 0.01.

Journal: Journal of Functional Foods

Article Title: Astragalin protects against lipopolysaccharide-triggered acute liver injury through suppression of necroptosis and inflammation and improvement of energy metabolism

doi: 10.1016/j.jff.2024.106298

Figure Lengend Snippet: Fig. 7. Astragalin (AST) eliminated lipopolysaccharide (LPS)-induced inflammatory response by inhibiting glycolysis enhancement mediated by the hypoxia- inducible factor-1α/pyruvate kinase M2 (HIF-1α/PKM2) signaling pathway. (A) LPS changed the concentrations of adenosine triphosphate (ATP) and lactic acid (LA) in liver and serum, and AST reversed these changes. Student’s t-test was performed, n = 4, * P < 0.05, ** P < 0.01, *** P < 0.001. (B) The activities of hexokinase (HK), pyruvate kinase (PK), and lactate dehydrogenase (LDH) in liver and serum were enhanced by LPS, whereas AST inhibited the enhancement. Student’s t-test was performed, n = 4, * P < 0.05, ** P < 0.01, *** P < 0.001. (C) Glycolysis-related genes, including HK2, LDHA, glucose-6-phosphate dehydrogenase X-linked (G6pdx), and glucose transporter 1 (Glut-1), were upregulated by LPS but AST prevented the upregulation. Similar results also showed in the expression of HIF-1α and PKM2 genes. Student’s t-test was performed, n = 4, * P < 0.05, ** P < 0.01, *** P < 0.001. (D-E) Expressions of HIF-1α and PKM2 proteins were increased by LPS treatment, while AST decreased their expression. Student’s t-test was performed, n = 3, * P < 0.05, ** P < 0.01.

Article Snippet: Adenosine triphosphate (ATP) content detection kit (#BC0305), lactic acid (LA) content assay kit (#BC2235), lactate dehydrogenase (LDH) activity assay kit (#BC0685), hexokinase (HK) activity assay kit (#BC0745), pyruvate kinase (PK) activity assay kit (#BC0545) were obtained from Beijing Solarbio Science & Technology Co., Ltd. Trizol (total RNA extraction reagent, #R0016), RIPA lysis buffer (strong, #P0013B), and BCA protein assay kit (#P0011) were provided by Beyotime Biotech Inc. Plus All-in-one 1st strand cDNA synthesis supermix (gDNA Purge, #E047-01B) and SYBR qPCR supermix plus (#E096-01B) were obtained from Novoprotein Scientific Inc. We got TNF-α polyclonal antibody (#17590–1-AP), TNFR1associated death domain protein (TRADD) polyclonal antibody (#15468–1-AP), HIF-1α polyclonal antibody (#20960–1-AP), PKM2, muscle2-specific monoclonal antibody (#60268–1-Ig), β-actin recombinant antibody (#81115–1-RR), and TANK-binding kinase 1 (TBK1) polyclonal antibody (#28397–1-AP) from Proteintech Group, Inc. Phospho-TBK1 (P-TBK1, Ser172) polyclonal antibody (#BD-PP1527) was provided by Biodragon Co. Ltd. Anti-phospho-RIP family of serinethreonine kinases (Ser166, P-RIPK1) polyclonal antibodies (#31122S), RIPK1 (D94C12) monoclonal antibody (#3493S), RIPK3 (D8J3L) monoclonal antibody (#15828S), and anti-phospho-RIPK3 (Thr231/ Ser232, E7S1R, #91702) were obtained from Cell Signaling Technology, Inc. Anti-phospho-MLKL (Ser345, P-MLKL) monoclonal antibody (#MABC1158) was provided by Merck Millipore Corporation, while MLKL polyclonal antibody (#PA5-71886) was provided by Thermo Fisher Scientific Inc. Bio-Rad Laboratories Co. Ltd provided CD68 monoclonal antibody (#MCA1957).

Techniques: Expressing

Fig. 8. The involved molecular mechanisms of astragalin (AST) in the treatment of lipopolysaccharide (LPS)-induced acute liver injury (ALI). LPS promotes the M1 type macrophage activation, causing inflammatory responses and the release of inflammatory cytokines, including tumor necrosis factor (TNF)-α. By binding to its receptor, TNF-α activated the receptor-interacting protein kinase 1 (RIPK1)/RIPK3/ mixed lineage kinase domain-like protein (MLKL) signal axis and mediated necroptosis. Meanwhile, LPS triggered the hypoxia-inducible factor-1α/pyruvate kinase M2 (HIF-1α/PKM2) signaling pathway, which mediates glycolysis-enhanced inflammatory response and oxidative stress. However, AST inhibited the transition of macrophages into M1 type, reduced the expression of pro-inflammatory cy tokines, which further decreased the activation of related signaling pathways, thereby preventing the liver from acute injury. This figure is created by Figdraw.

Journal: Journal of Functional Foods

Article Title: Astragalin protects against lipopolysaccharide-triggered acute liver injury through suppression of necroptosis and inflammation and improvement of energy metabolism

doi: 10.1016/j.jff.2024.106298

Figure Lengend Snippet: Fig. 8. The involved molecular mechanisms of astragalin (AST) in the treatment of lipopolysaccharide (LPS)-induced acute liver injury (ALI). LPS promotes the M1 type macrophage activation, causing inflammatory responses and the release of inflammatory cytokines, including tumor necrosis factor (TNF)-α. By binding to its receptor, TNF-α activated the receptor-interacting protein kinase 1 (RIPK1)/RIPK3/ mixed lineage kinase domain-like protein (MLKL) signal axis and mediated necroptosis. Meanwhile, LPS triggered the hypoxia-inducible factor-1α/pyruvate kinase M2 (HIF-1α/PKM2) signaling pathway, which mediates glycolysis-enhanced inflammatory response and oxidative stress. However, AST inhibited the transition of macrophages into M1 type, reduced the expression of pro-inflammatory cy tokines, which further decreased the activation of related signaling pathways, thereby preventing the liver from acute injury. This figure is created by Figdraw.

Article Snippet: Adenosine triphosphate (ATP) content detection kit (#BC0305), lactic acid (LA) content assay kit (#BC2235), lactate dehydrogenase (LDH) activity assay kit (#BC0685), hexokinase (HK) activity assay kit (#BC0745), pyruvate kinase (PK) activity assay kit (#BC0545) were obtained from Beijing Solarbio Science & Technology Co., Ltd. Trizol (total RNA extraction reagent, #R0016), RIPA lysis buffer (strong, #P0013B), and BCA protein assay kit (#P0011) were provided by Beyotime Biotech Inc. Plus All-in-one 1st strand cDNA synthesis supermix (gDNA Purge, #E047-01B) and SYBR qPCR supermix plus (#E096-01B) were obtained from Novoprotein Scientific Inc. We got TNF-α polyclonal antibody (#17590–1-AP), TNFR1associated death domain protein (TRADD) polyclonal antibody (#15468–1-AP), HIF-1α polyclonal antibody (#20960–1-AP), PKM2, muscle2-specific monoclonal antibody (#60268–1-Ig), β-actin recombinant antibody (#81115–1-RR), and TANK-binding kinase 1 (TBK1) polyclonal antibody (#28397–1-AP) from Proteintech Group, Inc. Phospho-TBK1 (P-TBK1, Ser172) polyclonal antibody (#BD-PP1527) was provided by Biodragon Co. Ltd. Anti-phospho-RIP family of serinethreonine kinases (Ser166, P-RIPK1) polyclonal antibodies (#31122S), RIPK1 (D94C12) monoclonal antibody (#3493S), RIPK3 (D8J3L) monoclonal antibody (#15828S), and anti-phospho-RIPK3 (Thr231/ Ser232, E7S1R, #91702) were obtained from Cell Signaling Technology, Inc. Anti-phospho-MLKL (Ser345, P-MLKL) monoclonal antibody (#MABC1158) was provided by Merck Millipore Corporation, while MLKL polyclonal antibody (#PA5-71886) was provided by Thermo Fisher Scientific Inc. Bio-Rad Laboratories Co. Ltd provided CD68 monoclonal antibody (#MCA1957).

Techniques: Activation Assay, Binding Assay, Expressing, Protein-Protein interactions

(A) Diagram of quadruple baculovirus transfer vector encoding influenza virus genes HA, NA, M1, and M2. This vector was transfected together with linearized baculovirus DNA into Sf9 insect cells to generate the quadruple recombinant (HA/Q28). Replacement of the HA with a gene encoding a full-length sequence of the VSV-G protein or chimera VSV-G/HA (see below) generated VSV-G/Q or VSV-G/HA/Q, respectively. (B) Structure of the VSV-G/HA chimera. The 11 aa of the cytoplasmic tail, 26 aa of the transmembrane domain, and 5 aa of the ectodomain of the influenza virus HA were fused in frame with the ectodomain of the VSV-G surface glycoprotein. This hybrid gene was used to replace the HA from the quadruple transfer vector (panel A) and generate the VSV-G/HA/Q gene.

Journal:

Article Title: Formation of Wild-Type and Chimeric Influenza Virus-Like Particles following Simultaneous Expression of Only Four Structural Proteins

doi: 10.1128/JVI.75.13.6154-6165.2001

Figure Lengend Snippet: (A) Diagram of quadruple baculovirus transfer vector encoding influenza virus genes HA, NA, M1, and M2. This vector was transfected together with linearized baculovirus DNA into Sf9 insect cells to generate the quadruple recombinant (HA/Q28). Replacement of the HA with a gene encoding a full-length sequence of the VSV-G protein or chimera VSV-G/HA (see below) generated VSV-G/Q or VSV-G/HA/Q, respectively. (B) Structure of the VSV-G/HA chimera. The 11 aa of the cytoplasmic tail, 26 aa of the transmembrane domain, and 5 aa of the ectodomain of the influenza virus HA were fused in frame with the ectodomain of the VSV-G surface glycoprotein. This hybrid gene was used to replace the HA from the quadruple transfer vector (panel A) and generate the VSV-G/HA/Q gene.

Article Snippet: The antibodies used in this work were obtained from the following sources: rat monoclonal anti-HA (clone 3F10), mouse monoclonal anti-HA (clone 12CA5), and mouse monoclonal anti-VSV-G (clone P5D4) were obtained from Roche Molecular Biochemicals (Indianapolis, Ind.); mouse monoclonal anti-M1 (clone GA2B) was from Serotec (Raleigh, N.C.); goat polyclonal anti-M1 was from Accurate Chemical & Scientific Corp. (Westbury, N.Y.); mouse monoclonal anti-NA (clone ST9D2) was from Argene Inc. (Massapequa, N.Y.); mouse monoclonal anti-M2 was from Mt.

Techniques: Plasmid Preparation, Transfection, Recombinant, Sequencing, Generated

Western blot analysis of the influenza virus and VSV-G proteins expressed in Sf9 cells infected by quadruple baculovirus recombinants (HA/Q28 or VSV-G/Q). (A) Expression of the influenza virus proteins HA, NA, and M2 in the cell pellet (lane 2) and culture supernatant (lane 3) was evaluated with a mixture of anti-HA, anti-M1, and anti-M2 monoclonal antibodies. Uninfected Sf9 cells and influenza virus A/Udorn-infected MDCK cells were used as controls (lanes 1 and 4, respectively). (B) Expression of VSV-G as well as influenza virus M1 and M2 proteins in Sf9 cells infected with VSV-G/Q (full-length G) was evaluated in cell pellets (lane 2) and culture supernatants (lane 3) by using a mixture of anti-G, anti-M1, and anti-M2 monoclonal antibodies. Uninfected Sf9 cells (lane 1), and VSV-infected BHK cells (lane 4) and influenza virus A/Udorn-infected MDCK cells (lane 5) were used as negative and positive controls, respectively.

Journal:

Article Title: Formation of Wild-Type and Chimeric Influenza Virus-Like Particles following Simultaneous Expression of Only Four Structural Proteins

doi: 10.1128/JVI.75.13.6154-6165.2001

Figure Lengend Snippet: Western blot analysis of the influenza virus and VSV-G proteins expressed in Sf9 cells infected by quadruple baculovirus recombinants (HA/Q28 or VSV-G/Q). (A) Expression of the influenza virus proteins HA, NA, and M2 in the cell pellet (lane 2) and culture supernatant (lane 3) was evaluated with a mixture of anti-HA, anti-M1, and anti-M2 monoclonal antibodies. Uninfected Sf9 cells and influenza virus A/Udorn-infected MDCK cells were used as controls (lanes 1 and 4, respectively). (B) Expression of VSV-G as well as influenza virus M1 and M2 proteins in Sf9 cells infected with VSV-G/Q (full-length G) was evaluated in cell pellets (lane 2) and culture supernatants (lane 3) by using a mixture of anti-G, anti-M1, and anti-M2 monoclonal antibodies. Uninfected Sf9 cells (lane 1), and VSV-infected BHK cells (lane 4) and influenza virus A/Udorn-infected MDCK cells (lane 5) were used as negative and positive controls, respectively.

Article Snippet: The antibodies used in this work were obtained from the following sources: rat monoclonal anti-HA (clone 3F10), mouse monoclonal anti-HA (clone 12CA5), and mouse monoclonal anti-VSV-G (clone P5D4) were obtained from Roche Molecular Biochemicals (Indianapolis, Ind.); mouse monoclonal anti-M1 (clone GA2B) was from Serotec (Raleigh, N.C.); goat polyclonal anti-M1 was from Accurate Chemical & Scientific Corp. (Westbury, N.Y.); mouse monoclonal anti-NA (clone ST9D2) was from Argene Inc. (Massapequa, N.Y.); mouse monoclonal anti-M2 was from Mt.

Techniques: Western Blot, Infection, Expressing

Immunofluorescence analysis of Sf9 cells infected with a quadruple baculovirus recombinant (HA/Q28). Expression of the influenza virus proteins HA, M1, or HA/M1 (A) and HA, NA, or HA/NA (B) was detected with a combination of anti-HA and anti-M1 or anti-HA and anti-NA antibodies as described in Materials and Methods. (C) M2 expression was examined with a rabbit anti-M2 peptide antibody. (D) Sf9 cells infected with VSV/Q chimeric baculovirus recombinant were used to evaluate expression of the VSV-G protein.

Journal:

Article Title: Formation of Wild-Type and Chimeric Influenza Virus-Like Particles following Simultaneous Expression of Only Four Structural Proteins

doi: 10.1128/JVI.75.13.6154-6165.2001

Figure Lengend Snippet: Immunofluorescence analysis of Sf9 cells infected with a quadruple baculovirus recombinant (HA/Q28). Expression of the influenza virus proteins HA, M1, or HA/M1 (A) and HA, NA, or HA/NA (B) was detected with a combination of anti-HA and anti-M1 or anti-HA and anti-NA antibodies as described in Materials and Methods. (C) M2 expression was examined with a rabbit anti-M2 peptide antibody. (D) Sf9 cells infected with VSV/Q chimeric baculovirus recombinant were used to evaluate expression of the VSV-G protein.

Article Snippet: The antibodies used in this work were obtained from the following sources: rat monoclonal anti-HA (clone 3F10), mouse monoclonal anti-HA (clone 12CA5), and mouse monoclonal anti-VSV-G (clone P5D4) were obtained from Roche Molecular Biochemicals (Indianapolis, Ind.); mouse monoclonal anti-M1 (clone GA2B) was from Serotec (Raleigh, N.C.); goat polyclonal anti-M1 was from Accurate Chemical & Scientific Corp. (Westbury, N.Y.); mouse monoclonal anti-NA (clone ST9D2) was from Argene Inc. (Massapequa, N.Y.); mouse monoclonal anti-M2 was from Mt.

Techniques: Immunofluorescence, Infection, Recombinant, Expressing

Analysis of VLP formation by iodixanol gradient centrifugation. Concentrated supernatants of HA/Q28- or VSV-G/Q recombinant-infected Sf9 cells were layered on top of an iodixanol density gradient and spun at 200,000 × g for 3.5 h. Fractions were collected from the bottom of the tube, resolved by SDS-PAGE, and transferred onto nitrocellulose. (A) Fractions from HA/Q28 were probed with a mixture of anti-HA and anti-M1 monoclonal antibodies. Lanes 1 to 8, fractions collected from top to bottom of the tube; lane 9, influenza virus A/Udorn-infected MDCK cells as control. (B) Fractions from VSV-G/Q were probed with a mixture of anti-VSV-G, anti-M1, and anti-M2 monoclonal antibodies. Lanes 1 to 8, fractions collected from the top to the bottom of the tube; lane 9, VSV-infected BHK cells and influenza virus-infected MDCK cells combined as control. (C) Concentrated supernatants of Sf9 cells infected with the M1 single baculovirus recombinant were purified as above and probed with anti-M1 antibody. Lanes 1 to 8, gradient fractions collected from the top to the bottom of the tube; lane 9, influenza virus-infected MDCK cells (control).

Journal:

Article Title: Formation of Wild-Type and Chimeric Influenza Virus-Like Particles following Simultaneous Expression of Only Four Structural Proteins

doi: 10.1128/JVI.75.13.6154-6165.2001

Figure Lengend Snippet: Analysis of VLP formation by iodixanol gradient centrifugation. Concentrated supernatants of HA/Q28- or VSV-G/Q recombinant-infected Sf9 cells were layered on top of an iodixanol density gradient and spun at 200,000 × g for 3.5 h. Fractions were collected from the bottom of the tube, resolved by SDS-PAGE, and transferred onto nitrocellulose. (A) Fractions from HA/Q28 were probed with a mixture of anti-HA and anti-M1 monoclonal antibodies. Lanes 1 to 8, fractions collected from top to bottom of the tube; lane 9, influenza virus A/Udorn-infected MDCK cells as control. (B) Fractions from VSV-G/Q were probed with a mixture of anti-VSV-G, anti-M1, and anti-M2 monoclonal antibodies. Lanes 1 to 8, fractions collected from the top to the bottom of the tube; lane 9, VSV-infected BHK cells and influenza virus-infected MDCK cells combined as control. (C) Concentrated supernatants of Sf9 cells infected with the M1 single baculovirus recombinant were purified as above and probed with anti-M1 antibody. Lanes 1 to 8, gradient fractions collected from the top to the bottom of the tube; lane 9, influenza virus-infected MDCK cells (control).

Article Snippet: The antibodies used in this work were obtained from the following sources: rat monoclonal anti-HA (clone 3F10), mouse monoclonal anti-HA (clone 12CA5), and mouse monoclonal anti-VSV-G (clone P5D4) were obtained from Roche Molecular Biochemicals (Indianapolis, Ind.); mouse monoclonal anti-M1 (clone GA2B) was from Serotec (Raleigh, N.C.); goat polyclonal anti-M1 was from Accurate Chemical & Scientific Corp. (Westbury, N.Y.); mouse monoclonal anti-NA (clone ST9D2) was from Argene Inc. (Massapequa, N.Y.); mouse monoclonal anti-M2 was from Mt.

Techniques: Gradient Centrifugation, Recombinant, Infection, SDS Page, Purification

(A) Comparative Western blot of M1 protein released from influenza virus A/Udorn-infected MDCK cells or from Sf9 cells infected with the quadruple recombinant HA/Q28 or the M1 single recombinant. MDCK cells (5 × 106) were infected with influenza virus A/Udorn (MOI, 1) for 24 h. Equivalent numbers of Sf9 cells were infected with either HA/Q28 or M1 single recombinant (MOI, 5) for 72 h. Supernatants were concentrated and particles were purified by iodixanol gradient centrifugation. Fractions 2 and 3 were pooled, and M1 protein was immunoprecipitated with anti-M1 monoclonal antibody twice to ensure complete recovery. Samples were resolved by SDS-PAGE, transferred to nitrocellulose, and probed with anti-M1 polyclonal antibody. Different amounts of each sample were analyzed to determine the linear range of the detecting color reaction. Lanes 1, 2, and 3 show the relative amount of M1 protein immunoprecipitated and detected by Western blotting from purified influenza virions, HA/Q 28, and M1 particles, respectively. We used NIH Image 1.54 to compare the relative amount of M1 protein detected in each sample. (B) Relative amounts of M1 and VSV-G proteins detected in culture supernatants of Sf9 cells infected with VSV-G/Q or VSV-G/HA/Q. Equivalent numbers of Sf9 cells (5 × 106) were infected with either VSV-G/Q or VSV-G/HA/Q quadruple recombinants (MOI, 5). Culture supernatants were concentrated, and the relative amounts of M1 and VSV-G or VSV-G/HA chimera were measured as described above. Lanes 1 and 2 show the M1 and VSV-G or VSV-G/HA chimeric proteins released from Sf9 cells infected with VSV-G/Q or VSV-G/HA/Q quadruple recombinants, respectively.

Journal:

Article Title: Formation of Wild-Type and Chimeric Influenza Virus-Like Particles following Simultaneous Expression of Only Four Structural Proteins

doi: 10.1128/JVI.75.13.6154-6165.2001

Figure Lengend Snippet: (A) Comparative Western blot of M1 protein released from influenza virus A/Udorn-infected MDCK cells or from Sf9 cells infected with the quadruple recombinant HA/Q28 or the M1 single recombinant. MDCK cells (5 × 106) were infected with influenza virus A/Udorn (MOI, 1) for 24 h. Equivalent numbers of Sf9 cells were infected with either HA/Q28 or M1 single recombinant (MOI, 5) for 72 h. Supernatants were concentrated and particles were purified by iodixanol gradient centrifugation. Fractions 2 and 3 were pooled, and M1 protein was immunoprecipitated with anti-M1 monoclonal antibody twice to ensure complete recovery. Samples were resolved by SDS-PAGE, transferred to nitrocellulose, and probed with anti-M1 polyclonal antibody. Different amounts of each sample were analyzed to determine the linear range of the detecting color reaction. Lanes 1, 2, and 3 show the relative amount of M1 protein immunoprecipitated and detected by Western blotting from purified influenza virions, HA/Q 28, and M1 particles, respectively. We used NIH Image 1.54 to compare the relative amount of M1 protein detected in each sample. (B) Relative amounts of M1 and VSV-G proteins detected in culture supernatants of Sf9 cells infected with VSV-G/Q or VSV-G/HA/Q. Equivalent numbers of Sf9 cells (5 × 106) were infected with either VSV-G/Q or VSV-G/HA/Q quadruple recombinants (MOI, 5). Culture supernatants were concentrated, and the relative amounts of M1 and VSV-G or VSV-G/HA chimera were measured as described above. Lanes 1 and 2 show the M1 and VSV-G or VSV-G/HA chimeric proteins released from Sf9 cells infected with VSV-G/Q or VSV-G/HA/Q quadruple recombinants, respectively.

Article Snippet: The antibodies used in this work were obtained from the following sources: rat monoclonal anti-HA (clone 3F10), mouse monoclonal anti-HA (clone 12CA5), and mouse monoclonal anti-VSV-G (clone P5D4) were obtained from Roche Molecular Biochemicals (Indianapolis, Ind.); mouse monoclonal anti-M1 (clone GA2B) was from Serotec (Raleigh, N.C.); goat polyclonal anti-M1 was from Accurate Chemical & Scientific Corp. (Westbury, N.Y.); mouse monoclonal anti-NA (clone ST9D2) was from Argene Inc. (Massapequa, N.Y.); mouse monoclonal anti-M2 was from Mt.

Techniques: Western Blot, Infection, Recombinant, Purification, Gradient Centrifugation, Immunoprecipitation, SDS Page

Deletion of Suv4-20h2 enhances influenza viral replication (A) WT mice were intratracheally infected with influenza virus (PR8 virus, 100 FFU), and lung tissues were sampled at the indicated time points. Histone was extracted from the lung tissues and was subjected to H4-tail proteomics analysis. Relative expression levels of H4K20me1, H4K20me2, and H4K20me3 to H4 were shown (n = 5 per group). (B–D) MEFs obtained from WT, Suv4-20h1 KO (h1 KO), Suv4-20h2 KO (h2 KO) and Suv4-20h1/Suv4-20h2 double KO (dKO) mice were infected with mock (Flu-) or influenza virus (MOI: 0.5) (Flu+). Cells were fixed and stained with either H4K20me1, H4K20me2 or H4K20me3 Abs, and Hoechst. Data are from three separate experiments. Representative staining is shown. Scale bars: 10 μm (B). Culture supernatant and cells were sampled at 8 hr after infection. Virus titer assessed by FFU in supernatant is shown (C, upper panel). NP mRNA expression of the cells is shown (C, lower panel). Data are from three separate experiments. ∗p < 0.05 compared to WT. Infected cells were fixed, and virus NP protein was stained (green). Nuclei were counterstained with Hoechst (blue). Representative staining is shown. Scale bars: 50 μm (D, left panel). The percentages of NP-positive in total of Hoechst positive are shown (D, right panel). ∗∗p < 0.01 compared to WT. (E) WT and dKO MEFs were infected with influenza virus (MOI: 0.5) and sampled at the indicated time point. Western blots of virus PB2, NP and M1 proteins, along with GAPDH are shown. Data were reproduced in three experiments. The data in (A, C and D) (right panel) are presented as means ± s.e.m.. Statistical analysis were carried out using analysis of variance with Bonferroni post t tests.

Journal: iScience

Article Title: Suv4-20h2 protects against influenza virus infection by suppression of chromatin loop formation

doi: 10.1016/j.isci.2021.102660

Figure Lengend Snippet: Deletion of Suv4-20h2 enhances influenza viral replication (A) WT mice were intratracheally infected with influenza virus (PR8 virus, 100 FFU), and lung tissues were sampled at the indicated time points. Histone was extracted from the lung tissues and was subjected to H4-tail proteomics analysis. Relative expression levels of H4K20me1, H4K20me2, and H4K20me3 to H4 were shown (n = 5 per group). (B–D) MEFs obtained from WT, Suv4-20h1 KO (h1 KO), Suv4-20h2 KO (h2 KO) and Suv4-20h1/Suv4-20h2 double KO (dKO) mice were infected with mock (Flu-) or influenza virus (MOI: 0.5) (Flu+). Cells were fixed and stained with either H4K20me1, H4K20me2 or H4K20me3 Abs, and Hoechst. Data are from three separate experiments. Representative staining is shown. Scale bars: 10 μm (B). Culture supernatant and cells were sampled at 8 hr after infection. Virus titer assessed by FFU in supernatant is shown (C, upper panel). NP mRNA expression of the cells is shown (C, lower panel). Data are from three separate experiments. ∗p < 0.05 compared to WT. Infected cells were fixed, and virus NP protein was stained (green). Nuclei were counterstained with Hoechst (blue). Representative staining is shown. Scale bars: 50 μm (D, left panel). The percentages of NP-positive in total of Hoechst positive are shown (D, right panel). ∗∗p < 0.01 compared to WT. (E) WT and dKO MEFs were infected with influenza virus (MOI: 0.5) and sampled at the indicated time point. Western blots of virus PB2, NP and M1 proteins, along with GAPDH are shown. Data were reproduced in three experiments. The data in (A, C and D) (right panel) are presented as means ± s.e.m.. Statistical analysis were carried out using analysis of variance with Bonferroni post t tests.

Article Snippet: Human SET domain of SUV4-20h2 (2-281) (Active motif) was pre-incubated with virus NP or M1 protein (SinoBiological) (0, 0.05, 0.5, or 1.0 μg) at room temperature for 1 hr and then incubated with polynucleosomes (Active motif) (1.0 μg) for additional 3 hr.

Techniques: Infection, Expressing, Staining, Western Blot

Suv4-20h2 interacts with virus and host nuclear proteins (A) dKO cells transduced with EGFP-tagged empty vector (EV), Suv4-20h2 full length (h2-FL), SET domain (h2-SET) or Clamp domain (h2-Clamp; left panel) were infected with influenza virus (MOI: 0.5) for 8 hr h2-EGFP and virus NP protein were co-stained. Nuclei were counterstained with Hoechst. The percentages of NP-positive in total of the h2 EGFP-positive are shown. Data are from three separate experiments. ∗∗P < 0.01 between the groups. (B) WT, dKO, and dKO MEFs overexpressed with Suv4-20h2 were infected with influenza virus (MOI: 0.5). Culture supernatant and cells were sampled at 8 hr after infection. Virus titer of FFU in supernatant is shown (B, left panel). NP mRNA expression of the cells is shown (B, right panel). Data are from three separate experiments. ∗p < 0.05, ∗∗p < 0.01 between the groups. (C) dKO MEFs transduced with AM-tagged Suv4-20h2 were infected with influenza virus (MOI: 0.5) for 8 hr. Nuclear extracts were prepared from the cells, and were immunoprecipitated (IP) with AM Ab, and IP product was subjected to a stable isotope labeling (SILAC) & LC-MS/MS analysis. Virus NP and PB2 proteins were identified in this analysis. (D) Suv4-20h2 Flag knock-in mouse embryonic stem (mES) cells were mock (Flu-) or infected with influenza virus (MOI: 0.5) (Flu+) for 8 hr. Nuclear extracts were prepared from the cells, and were immunoprecipitated with Flag Ab. Western blotting shows that virus NP and PB2 proteins were co-precipitated with Suv4-20h2. Data were reproduced in three experiments. (E) Purified recombinant viral NP and PB2 proteins and GST-fusion proteins to full length (FL), SET, and Clamp domain of Suv4-20h2, were used in a pull-down assay. Virus NP and PB2 proteins interacted with FL and SET domain of Suv4-20h2. Data were reproduced in three experiments. (F) Human SET domain of SUV4-20H2 (2-281) was pre-incubated with virus NP protein (0, 0.05, 0.5, or 1.0 μg) (left panel) or M1 protein (0, 0.05, 0.5, or 1.0 μg) (right panel) at room temperature for 1hr and then incubated with polynucleosomes (1.0 μg) for additional 3hr. Protein samples were subjected to Western blot analysis using specific antibodies. H4K20me3 was suppressed in a dose-dependent manner by NP protein. Data were reproduced in three experiments. The data in (A and B) are presented as means ± s.e.m.. Statistical analysis were carried out using analysis of variance with Bonferroni post t tests.

Journal: iScience

Article Title: Suv4-20h2 protects against influenza virus infection by suppression of chromatin loop formation

doi: 10.1016/j.isci.2021.102660

Figure Lengend Snippet: Suv4-20h2 interacts with virus and host nuclear proteins (A) dKO cells transduced with EGFP-tagged empty vector (EV), Suv4-20h2 full length (h2-FL), SET domain (h2-SET) or Clamp domain (h2-Clamp; left panel) were infected with influenza virus (MOI: 0.5) for 8 hr h2-EGFP and virus NP protein were co-stained. Nuclei were counterstained with Hoechst. The percentages of NP-positive in total of the h2 EGFP-positive are shown. Data are from three separate experiments. ∗∗P < 0.01 between the groups. (B) WT, dKO, and dKO MEFs overexpressed with Suv4-20h2 were infected with influenza virus (MOI: 0.5). Culture supernatant and cells were sampled at 8 hr after infection. Virus titer of FFU in supernatant is shown (B, left panel). NP mRNA expression of the cells is shown (B, right panel). Data are from three separate experiments. ∗p < 0.05, ∗∗p < 0.01 between the groups. (C) dKO MEFs transduced with AM-tagged Suv4-20h2 were infected with influenza virus (MOI: 0.5) for 8 hr. Nuclear extracts were prepared from the cells, and were immunoprecipitated (IP) with AM Ab, and IP product was subjected to a stable isotope labeling (SILAC) & LC-MS/MS analysis. Virus NP and PB2 proteins were identified in this analysis. (D) Suv4-20h2 Flag knock-in mouse embryonic stem (mES) cells were mock (Flu-) or infected with influenza virus (MOI: 0.5) (Flu+) for 8 hr. Nuclear extracts were prepared from the cells, and were immunoprecipitated with Flag Ab. Western blotting shows that virus NP and PB2 proteins were co-precipitated with Suv4-20h2. Data were reproduced in three experiments. (E) Purified recombinant viral NP and PB2 proteins and GST-fusion proteins to full length (FL), SET, and Clamp domain of Suv4-20h2, were used in a pull-down assay. Virus NP and PB2 proteins interacted with FL and SET domain of Suv4-20h2. Data were reproduced in three experiments. (F) Human SET domain of SUV4-20H2 (2-281) was pre-incubated with virus NP protein (0, 0.05, 0.5, or 1.0 μg) (left panel) or M1 protein (0, 0.05, 0.5, or 1.0 μg) (right panel) at room temperature for 1hr and then incubated with polynucleosomes (1.0 μg) for additional 3hr. Protein samples were subjected to Western blot analysis using specific antibodies. H4K20me3 was suppressed in a dose-dependent manner by NP protein. Data were reproduced in three experiments. The data in (A and B) are presented as means ± s.e.m.. Statistical analysis were carried out using analysis of variance with Bonferroni post t tests.

Article Snippet: Human SET domain of SUV4-20h2 (2-281) (Active motif) was pre-incubated with virus NP or M1 protein (SinoBiological) (0, 0.05, 0.5, or 1.0 μg) at room temperature for 1 hr and then incubated with polynucleosomes (Active motif) (1.0 μg) for additional 3 hr.

Techniques: Transduction, Plasmid Preparation, Infection, Staining, Expressing, Immunoprecipitation, Labeling, Liquid Chromatography with Mass Spectroscopy, Knock-In, Western Blot, Purification, Recombinant, Pull Down Assay, Incubation

Journal: iScience

Article Title: Suv4-20h2 protects against influenza virus infection by suppression of chromatin loop formation

doi: 10.1016/j.isci.2021.102660

Figure Lengend Snippet:

Article Snippet: Human SET domain of SUV4-20h2 (2-281) (Active motif) was pre-incubated with virus NP or M1 protein (SinoBiological) (0, 0.05, 0.5, or 1.0 μg) at room temperature for 1 hr and then incubated with polynucleosomes (Active motif) (1.0 μg) for additional 3 hr.

Techniques: Recombinant, Transfection, Protease Inhibitor, Western Blot, Purification, Clone Assay, Fractionation, Sequencing, Software

Identification of the ECM-related genes associated with patients survival. Notes . (a) Univariate Cox regression analysis was utilized to identify the prognosis-related genes with P < 0.05; (b) LASSO regression analysis; (c) multivariate cox regression analysis based on the genes screened by LASSO regression; (d–i) clinical correlation of IBSP, LINGO4, COL26A1, MMP12, KLK4, RTBDN, TENM1, GDF15, and RUNX1.

Journal: Genetics Research

Article Title: Impact of Extracellular Matrix-Related Genes on the Tumor Microenvironment and Prognostic Indicators in Esophageal Cancer: A Comprehensive Analytical Study

doi: 10.1155/2024/3577395

Figure Lengend Snippet: Identification of the ECM-related genes associated with patients survival. Notes . (a) Univariate Cox regression analysis was utilized to identify the prognosis-related genes with P < 0.05; (b) LASSO regression analysis; (c) multivariate cox regression analysis based on the genes screened by LASSO regression; (d–i) clinical correlation of IBSP, LINGO4, COL26A1, MMP12, KLK4, RTBDN, TENM1, GDF15, and RUNX1.

Article Snippet: Equal amounts of protein were loaded onto SDS-PAGE gels, transferred to PVDF membranes, and probed with antibodies specific to TENM1 (proteintech, 21696-1-AP, 1 : 1500) and GAPDH (proteintech, 60004-1-Ig, 1 : 10000).

Techniques:

Expression level of TENM1 in EC cells. Notes . (a) The mRNA level of TENM1 in EC and normal cells; (b) the protein level of TENM1 in EC and normal cells.

Journal: Genetics Research

Article Title: Impact of Extracellular Matrix-Related Genes on the Tumor Microenvironment and Prognostic Indicators in Esophageal Cancer: A Comprehensive Analytical Study

doi: 10.1155/2024/3577395

Figure Lengend Snippet: Expression level of TENM1 in EC cells. Notes . (a) The mRNA level of TENM1 in EC and normal cells; (b) the protein level of TENM1 in EC and normal cells.

Article Snippet: Equal amounts of protein were loaded onto SDS-PAGE gels, transferred to PVDF membranes, and probed with antibodies specific to TENM1 (proteintech, 21696-1-AP, 1 : 1500) and GAPDH (proteintech, 60004-1-Ig, 1 : 10000).

Techniques: Expressing