Journal: Pharmaceuticals

Article Title: Expression Analysis of α5 Integrin Subunit Reveals Its Upregulation as a Negative Prognostic Biomarker for Glioblastoma

doi: 10.3390/ph14090882

Figure Lengend Snippet: Clinicopathological features of the 95 GBM (glioblastoma) patients, comparing low integrin α5 and high integrin α5 groups according to median expression level (median of MMFI) as the cut-off threshold.

Article Snippet: Blots were probed with antibodies to α5 integrin (H104, Santa Cruz Biotechnology, Heidelberg, Germany) and to glyceraldehyde 3-phosphate dehydrogenase (GAPDH; Merck-Millipore, Molsheim, France), the latter used as the loading control for the tissue lysate samples.

Techniques: Expressing, Methylation

Journal: Pharmaceuticals

Article Title: Expression Analysis of α5 Integrin Subunit Reveals Its Upregulation as a Negative Prognostic Biomarker for Glioblastoma

doi: 10.3390/ph14090882

Figure Lengend Snippet: Immunofluorescence ( a ) and Western blot ( b ) on GBM-PDX tumor TC7 and TC22 presenting high and low levels of α5 integrin, respectively. In immunofluorescence, detection of integrin α5 (in red) was realized with AB1928 antibody followed by a secondary antibody coupled to Alexa Fluor ® 647. DAPI staining is shown in blue. One representative image per condition is shown (magnification ×63). In Western blot, detection of integrin α5 was realized in 3 xenografts from 3 different mice with H104 antibody. Anti-GAPDH antibody was used as a loading control antibody.

Article Snippet: Blots were probed with antibodies to α5 integrin (H104, Santa Cruz Biotechnology, Heidelberg, Germany) and to glyceraldehyde 3-phosphate dehydrogenase (GAPDH; Merck-Millipore, Molsheim, France), the latter used as the loading control for the tissue lysate samples.

Techniques: Immunofluorescence, Western Blot, Staining, Control

Journal: Pharmaceuticals

Article Title: Expression Analysis of α5 Integrin Subunit Reveals Its Upregulation as a Negative Prognostic Biomarker for Glioblastoma

doi: 10.3390/ph14090882

Figure Lengend Snippet: Quantification of integrin α5 protein expression level in glioblastoma. ( a ) Representative cases of low and high integrin α5 immunostaining (magnification ×40) and ( b ) distribution of cumulative data for integrin α5 expression level (MMFI expressed as arbitrary units/A.U.) in glioblastoma samples. The median of MMFI of the all cohort (275 A.U.) was used as a cut-off to distinguish 2 groups characterized by low and high integrin α5 expression levels.

Article Snippet: Blots were probed with antibodies to α5 integrin (H104, Santa Cruz Biotechnology, Heidelberg, Germany) and to glyceraldehyde 3-phosphate dehydrogenase (GAPDH; Merck-Millipore, Molsheim, France), the latter used as the loading control for the tissue lysate samples.

Techniques: Expressing, Immunostaining

Journal: Pharmaceuticals

Article Title: Expression Analysis of α5 Integrin Subunit Reveals Its Upregulation as a Negative Prognostic Biomarker for Glioblastoma

doi: 10.3390/ph14090882

Figure Lengend Snippet: Univariate analysis of factors associated with survival and progression.

Article Snippet: Blots were probed with antibodies to α5 integrin (H104, Santa Cruz Biotechnology, Heidelberg, Germany) and to glyceraldehyde 3-phosphate dehydrogenase (GAPDH; Merck-Millipore, Molsheim, France), the latter used as the loading control for the tissue lysate samples.

Techniques: Methylation, Expressing

Journal: Pharmaceuticals

Article Title: Expression Analysis of α5 Integrin Subunit Reveals Its Upregulation as a Negative Prognostic Biomarker for Glioblastoma

doi: 10.3390/ph14090882

Figure Lengend Snippet: Multivariate Cox regression analysis for survival.

Article Snippet: Blots were probed with antibodies to α5 integrin (H104, Santa Cruz Biotechnology, Heidelberg, Germany) and to glyceraldehyde 3-phosphate dehydrogenase (GAPDH; Merck-Millipore, Molsheim, France), the latter used as the loading control for the tissue lysate samples.

Techniques: Methylation

Journal: Pharmaceuticals

Article Title: Expression Analysis of α5 Integrin Subunit Reveals Its Upregulation as a Negative Prognostic Biomarker for Glioblastoma

doi: 10.3390/ph14090882

Figure Lengend Snippet: Overall survival (OS) estimated according to the expression level of integrin α5 and the completion of the Stupp protocol. Kaplan–Meier survival analysis for high and low expression levels of integrin α5 in the all cohort ( a ) in patients treated by the Stupp protocol ( b ) and in patients who completed the Stupp protocol ( c ); HR: hazard ratio in univariate analysis; HR adj : adjusted hazard ratio in multivariate analysis; CI: confidence interval. Survivals were compared using the log-rank (Mantel–Cox) test. p -values < 0.05 were considered as significant.

Article Snippet: Blots were probed with antibodies to α5 integrin (H104, Santa Cruz Biotechnology, Heidelberg, Germany) and to glyceraldehyde 3-phosphate dehydrogenase (GAPDH; Merck-Millipore, Molsheim, France), the latter used as the loading control for the tissue lysate samples.

Techniques: Expressing

Journal: Virologica Sinica

Article Title: African Swine Fever Virus MGF360-12L Inhibits Type I Interferon Production by Blocking the Interaction of Importin α and NF-κB Signaling Pathway

doi: 10.1007/s12250-020-00304-4

Figure Lengend Snippet: MGF360 - 12L may interact with KPNA2, KPNA3 and KPNA4 and disrupt the interaction between p65 and KPNA2, KPNA3, KPNA4. A The plasmid encoding MGF360 - 12L or empty vector (EV) were co-transfected into HeLa cells with plasmids EGFP-C1-GST-2VSV40T. After transfection 36 h, cells were fixed and stained with DAPI (blue), EGFP (green), anti-Flag (red) antibody and examined by confocal microscope. Magnification, 40×. B HeLa cells were transfected with ASFV pFlag-12L plasmid or empty vector. At 24 h and 48 h after transfection, collection cells, and the expression levels of endogenous KPNA1, KPNA2, KPNA3, KPNA4, KPNA5, KPNA6, and KPNB1 were analyzed by Western blotting. C Schematic diagram of NSL-deleted 12L . D HeLa cells were co-transfected with pFlag-12L or pFlag-Δ12L plasmid and the pMyc-KPNA2, pMyc-KPNA3 or pMyc-KPNA4. At 36 h after transfection, immunoprecipitation (IP) was performed with whole-cell lysates by using anti-Flag antibody. E HeLa cells were co-transfected with the pMyc-KPNA2, pMyc-KPNA3 or pMyc-KPNA4 and pFlag-12L plasmid or empty vector. At 24 h after transfection, cells were treated with TNFα (5 µg/mL) for 30 min and collected. Immunoprecipitation was performed with KPNA2, KPNA3 or KPNA4 antibodies, and Western blotting was performed with antibodies against KPNA2, KPNA3 or KPNA4, Flag and p65.

Article Snippet: The membranes were incubated for 2 h at room temperature in blocking buffer (20 mmol/L Tris-HCl, 137 mmol/L NaCl, pH 8.0, containing 0.1% Tween and 5% non-fat dry milk) and probed with antibodies against KPNA1 (18137-1-AP, Proteintech, China), KPNA2 (10819-1-AP, Proteintech, China), KPNA3 (abs118425, Absin, China), KPNA4 (12463-1-AP, Proteintech, China), KPNA5(13963-1-AP, Proteintech, China), KPNA6 (12366-2-AP, Proteintech, China), KPNB1 (10077-1-AP, Proteintech, China), Flag (20543-1-AP, Proteintech, China), Myc (60003-2-lg, Proteintech, China), p65 (10745-1-AP, Proteintech, China), p50 (14220-1-AP, Proteintech, China), IκBα (10268-1-AP, Proteintech, China), 1:2000 dilution; Histone H3 (1: 10000, GB13102-1, Servicebio, China), GAPDH (1: 5000, GB12002, Servicebio, China) antibodies at 4 °C overnight.

Techniques: Plasmid Preparation, Transfection, Staining, Microscopy, Expressing, Western Blot, Immunoprecipitation