glyceraldehyde 3 phosphate dehydrogenase gapdh Proteintech Search Results


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  • 95
    Millipore glyceraldehyde 3 phosphate dehydrogenase
    Glyceraldehyde 3 Phosphate Dehydrogenase, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 1570 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech glyceraldehyde 3 phosphate dehydrogenase gapdh
    Effects of DACT1 on β-catenin in myocardial cells. Effect of DACT1 overexpression on β-catenin concentration A) and distribution B) in H9C2 and HL-1 cells. C) Correlation between DACT1 and β-catenin expression in the myocardial tissues of patients with valvular heart disease. D) Colocation of DACT1 and β-catenin in the myocardial cells. H9C2 and HL-1 cells were overexpressed with the two isoforms of the DACT1 coding sequence, and then immunofluorescence was used to detect the DACT1 (DACT1-V1 or DACT1-V2) (OriGene) and β-catenin (CST) location. The secondary antibody Alexa Fluor 647 was used for DACT1 detection and Alexa Fluor 568 for β-catenin. The ZEN (Zeiss) software was used for further analysis. Alexa Fluor 647 was represented by the green color and Alexa Fluor 568 by the red color, while channel 488 was closed to avoid interference. DACT1=dishevelled binding antagonist of beta catenin 1; DAPI=2-(4-amidinophenyl)-6-indolecarbamidine dihydrochloride; <t>GAPDH=glyceraldehyde-3-phosphate</t> dehydrogenase
    Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Proteintech, used in various techniques. Bioz Stars score: 98/100, based on 166 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech glyceraldehyde 3 phosphate dehydrogenase gapdh antibodies
    Effect of lentivirus-mediated small hairpin RNA (shRNA) (S1) silencing on immature colon carcinoma transcript-1 (ICT1) expression in U251 cells. Notes: ( A ) Expression patterns of ICT1 in four glioblastoma multiforme (GBM) cell lines U251, U87, U373, and A172 determined by Western-blot analysis. ( B ) Green fluorescence protein (GFP) expression recorded under a fluorescence microscope. The representative pictures shown are from one of three independent experiments. ( C ) The messenger RNA levels of ICT1 in non-treated, constructed lentiviruses containing non-silencing small hairpin RNA (Lv-shCon)-treated and Lv-shICT1 (S1)-treated cells determined by RT-qPCR analysis. ( D ) The protein levels of ICT1 in non-treated, Lv-shCon-treated, and Lv-shICT1 (S1)-treated cells determined by Western-blot analysis. Scale bar in ( B ): 100 μm. Data are presented as mean ± standard deviation of three independent experiments performed in triplicate. *** P μ0.01 versus Lv-shCon. Abbreviations: con, control; <t>GAPDH,</t> glyceraldehyde 3-phosphate dehydrogenase.
    Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh Antibodies, supplied by Proteintech, used in various techniques. Bioz Stars score: 83/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech glyceraldehyde 3 phosphate dehydrogenase
    Effect of lentivirus-mediated small hairpin RNA (shRNA) (S1) silencing on immature colon carcinoma transcript-1 (ICT1) expression in U251 cells. Notes: ( A ) Expression patterns of ICT1 in four glioblastoma multiforme (GBM) cell lines U251, U87, U373, and A172 determined by Western-blot analysis. ( B ) Green fluorescence protein (GFP) expression recorded under a fluorescence microscope. The representative pictures shown are from one of three independent experiments. ( C ) The messenger RNA levels of ICT1 in non-treated, constructed lentiviruses containing non-silencing small hairpin RNA (Lv-shCon)-treated and Lv-shICT1 (S1)-treated cells determined by RT-qPCR analysis. ( D ) The protein levels of ICT1 in non-treated, Lv-shCon-treated, and Lv-shICT1 (S1)-treated cells determined by Western-blot analysis. Scale bar in ( B ): 100 μm. Data are presented as mean ± standard deviation of three independent experiments performed in triplicate. *** P μ0.01 versus Lv-shCon. Abbreviations: con, control; <t>GAPDH,</t> glyceraldehyde 3-phosphate dehydrogenase.
    Glyceraldehyde 3 Phosphate Dehydrogenase, supplied by Proteintech, used in various techniques. Bioz Stars score: 99/100, based on 73 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech mouse anti glyceraldehyde 3 phosphate dehydrogenase gapdh
    Effect of lentivirus-mediated small hairpin RNA (shRNA) (S1) silencing on immature colon carcinoma transcript-1 (ICT1) expression in U251 cells. Notes: ( A ) Expression patterns of ICT1 in four glioblastoma multiforme (GBM) cell lines U251, U87, U373, and A172 determined by Western-blot analysis. ( B ) Green fluorescence protein (GFP) expression recorded under a fluorescence microscope. The representative pictures shown are from one of three independent experiments. ( C ) The messenger RNA levels of ICT1 in non-treated, constructed lentiviruses containing non-silencing small hairpin RNA (Lv-shCon)-treated and Lv-shICT1 (S1)-treated cells determined by RT-qPCR analysis. ( D ) The protein levels of ICT1 in non-treated, Lv-shCon-treated, and Lv-shICT1 (S1)-treated cells determined by Western-blot analysis. Scale bar in ( B ): 100 μm. Data are presented as mean ± standard deviation of three independent experiments performed in triplicate. *** P μ0.01 versus Lv-shCon. Abbreviations: con, control; <t>GAPDH,</t> glyceraldehyde 3-phosphate dehydrogenase.
    Mouse Anti Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Proteintech, used in various techniques. Bioz Stars score: 99/100, based on 34 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech rabbit anti glyceraldehyde 3 phosphate dehydrogenase gapdh
    Effect of lentivirus-mediated small hairpin RNA (shRNA) (S1) silencing on immature colon carcinoma transcript-1 (ICT1) expression in U251 cells. Notes: ( A ) Expression patterns of ICT1 in four glioblastoma multiforme (GBM) cell lines U251, U87, U373, and A172 determined by Western-blot analysis. ( B ) Green fluorescence protein (GFP) expression recorded under a fluorescence microscope. The representative pictures shown are from one of three independent experiments. ( C ) The messenger RNA levels of ICT1 in non-treated, constructed lentiviruses containing non-silencing small hairpin RNA (Lv-shCon)-treated and Lv-shICT1 (S1)-treated cells determined by RT-qPCR analysis. ( D ) The protein levels of ICT1 in non-treated, Lv-shCon-treated, and Lv-shICT1 (S1)-treated cells determined by Western-blot analysis. Scale bar in ( B ): 100 μm. Data are presented as mean ± standard deviation of three independent experiments performed in triplicate. *** P μ0.01 versus Lv-shCon. Abbreviations: con, control; <t>GAPDH,</t> glyceraldehyde 3-phosphate dehydrogenase.
    Rabbit Anti Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Proteintech, used in various techniques. Bioz Stars score: 92/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech affinity purified anti glyceraldehyde 3 phosphate dehydrogenase gapdh antibody
    Effect of lentivirus-mediated small hairpin RNA (shRNA) (S1) silencing on immature colon carcinoma transcript-1 (ICT1) expression in U251 cells. Notes: ( A ) Expression patterns of ICT1 in four glioblastoma multiforme (GBM) cell lines U251, U87, U373, and A172 determined by Western-blot analysis. ( B ) Green fluorescence protein (GFP) expression recorded under a fluorescence microscope. The representative pictures shown are from one of three independent experiments. ( C ) The messenger RNA levels of ICT1 in non-treated, constructed lentiviruses containing non-silencing small hairpin RNA (Lv-shCon)-treated and Lv-shICT1 (S1)-treated cells determined by RT-qPCR analysis. ( D ) The protein levels of ICT1 in non-treated, Lv-shCon-treated, and Lv-shICT1 (S1)-treated cells determined by Western-blot analysis. Scale bar in ( B ): 100 μm. Data are presented as mean ± standard deviation of three independent experiments performed in triplicate. *** P μ0.01 versus Lv-shCon. Abbreviations: con, control; <t>GAPDH,</t> glyceraldehyde 3-phosphate dehydrogenase.
    Affinity Purified Anti Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 98/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech glyceraldehyde 3 phosphate dehydrogenase gapdh primary antibody
    Effect of lentivirus-mediated small hairpin RNA (shRNA) (S1) silencing on immature colon carcinoma transcript-1 (ICT1) expression in U251 cells. Notes: ( A ) Expression patterns of ICT1 in four glioblastoma multiforme (GBM) cell lines U251, U87, U373, and A172 determined by Western-blot analysis. ( B ) Green fluorescence protein (GFP) expression recorded under a fluorescence microscope. The representative pictures shown are from one of three independent experiments. ( C ) The messenger RNA levels of ICT1 in non-treated, constructed lentiviruses containing non-silencing small hairpin RNA (Lv-shCon)-treated and Lv-shICT1 (S1)-treated cells determined by RT-qPCR analysis. ( D ) The protein levels of ICT1 in non-treated, Lv-shCon-treated, and Lv-shICT1 (S1)-treated cells determined by Western-blot analysis. Scale bar in ( B ): 100 μm. Data are presented as mean ± standard deviation of three independent experiments performed in triplicate. *** P μ0.01 versus Lv-shCon. Abbreviations: con, control; <t>GAPDH,</t> glyceraldehyde 3-phosphate dehydrogenase.
    Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh Primary Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 99/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech rabbit gapdh polyclonal
    Effect of lentivirus-mediated small hairpin RNA (shRNA) (S1) silencing on immature colon carcinoma transcript-1 (ICT1) expression in U251 cells. Notes: ( A ) Expression patterns of ICT1 in four glioblastoma multiforme (GBM) cell lines U251, U87, U373, and A172 determined by Western-blot analysis. ( B ) Green fluorescence protein (GFP) expression recorded under a fluorescence microscope. The representative pictures shown are from one of three independent experiments. ( C ) The messenger RNA levels of ICT1 in non-treated, constructed lentiviruses containing non-silencing small hairpin RNA (Lv-shCon)-treated and Lv-shICT1 (S1)-treated cells determined by RT-qPCR analysis. ( D ) The protein levels of ICT1 in non-treated, Lv-shCon-treated, and Lv-shICT1 (S1)-treated cells determined by Western-blot analysis. Scale bar in ( B ): 100 μm. Data are presented as mean ± standard deviation of three independent experiments performed in triplicate. *** P μ0.01 versus Lv-shCon. Abbreviations: con, control; <t>GAPDH,</t> glyceraldehyde 3-phosphate dehydrogenase.
    Rabbit Gapdh Polyclonal, supplied by Proteintech, used in various techniques. Bioz Stars score: 90/100, based on 93 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech mouse anti glyceraldehyde 3 phosphate dehydrogenase
    Effect of lentivirus-mediated small hairpin RNA (shRNA) (S1) silencing on immature colon carcinoma transcript-1 (ICT1) expression in U251 cells. Notes: ( A ) Expression patterns of ICT1 in four glioblastoma multiforme (GBM) cell lines U251, U87, U373, and A172 determined by Western-blot analysis. ( B ) Green fluorescence protein (GFP) expression recorded under a fluorescence microscope. The representative pictures shown are from one of three independent experiments. ( C ) The messenger RNA levels of ICT1 in non-treated, constructed lentiviruses containing non-silencing small hairpin RNA (Lv-shCon)-treated and Lv-shICT1 (S1)-treated cells determined by RT-qPCR analysis. ( D ) The protein levels of ICT1 in non-treated, Lv-shCon-treated, and Lv-shICT1 (S1)-treated cells determined by Western-blot analysis. Scale bar in ( B ): 100 μm. Data are presented as mean ± standard deviation of three independent experiments performed in triplicate. *** P μ0.01 versus Lv-shCon. Abbreviations: con, control; <t>GAPDH,</t> glyceraldehyde 3-phosphate dehydrogenase.
    Mouse Anti Glyceraldehyde 3 Phosphate Dehydrogenase, supplied by Proteintech, used in various techniques. Bioz Stars score: 99/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech anti horseradish peroxidase hrp conjugated glyceraldehyde 3 phosphate dehydrogenase gapdh
    ISL induces cell cycle arrest in human endometrial cancer cells Cells were plated in 100 mm dishes at a density of 1 × 10 6 cells/plate in media supplemented with 10% FBS and allowed to adhere. The morphology of cells after treatment with vehicle or ISL (10, 25, 50 μΜ) in media containing 1% FBS for 48 h. ( A ) Ishikawa and ( B ) HEC-1A cells were stained with propidium iodide (PI), and cell cycle distribution was analyzed by flow cytometry. The vertical axis represents the number of cells, and the horizontal axis represents the intensity of PI staining. The cell cycle distribution is shown as a bar graph. The vertical numbers represent the cell population percentage in cell cycles S, G2, and G1phase, and the horizontal numbers represent the concentration of ISL. ( C ) Ishikawa and ( D ) HEC-1A cell lysates were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and analyzed by western blot with the indicated antibodies. Glyceraldehyde 3-phosphate dehydrogenase <t>(GAPDH)</t> was used as a loading control. The values of the band intensity represent the densitometric estimation of each band normalized to GAPDH.
    Anti Horseradish Peroxidase Hrp Conjugated Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Proteintech, used in various techniques. Bioz Stars score: 81/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech rabbit anti glyceraldehyde 3 phosphate dehydrogenase
    ISL induces cell cycle arrest in human endometrial cancer cells Cells were plated in 100 mm dishes at a density of 1 × 10 6 cells/plate in media supplemented with 10% FBS and allowed to adhere. The morphology of cells after treatment with vehicle or ISL (10, 25, 50 μΜ) in media containing 1% FBS for 48 h. ( A ) Ishikawa and ( B ) HEC-1A cells were stained with propidium iodide (PI), and cell cycle distribution was analyzed by flow cytometry. The vertical axis represents the number of cells, and the horizontal axis represents the intensity of PI staining. The cell cycle distribution is shown as a bar graph. The vertical numbers represent the cell population percentage in cell cycles S, G2, and G1phase, and the horizontal numbers represent the concentration of ISL. ( C ) Ishikawa and ( D ) HEC-1A cell lysates were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and analyzed by western blot with the indicated antibodies. Glyceraldehyde 3-phosphate dehydrogenase <t>(GAPDH)</t> was used as a loading control. The values of the band intensity represent the densitometric estimation of each band normalized to GAPDH.
    Rabbit Anti Glyceraldehyde 3 Phosphate Dehydrogenase, supplied by Proteintech, used in various techniques. Bioz Stars score: 99/100, based on 46 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech horseradish peroxidase hrp conjugated gapdh glyceraldehyde 3 phosphate dehydrogenase monoclonal antibody
    ISL induces cell cycle arrest in human endometrial cancer cells Cells were plated in 100 mm dishes at a density of 1 × 10 6 cells/plate in media supplemented with 10% FBS and allowed to adhere. The morphology of cells after treatment with vehicle or ISL (10, 25, 50 μΜ) in media containing 1% FBS for 48 h. ( A ) Ishikawa and ( B ) HEC-1A cells were stained with propidium iodide (PI), and cell cycle distribution was analyzed by flow cytometry. The vertical axis represents the number of cells, and the horizontal axis represents the intensity of PI staining. The cell cycle distribution is shown as a bar graph. The vertical numbers represent the cell population percentage in cell cycles S, G2, and G1phase, and the horizontal numbers represent the concentration of ISL. ( C ) Ishikawa and ( D ) HEC-1A cell lysates were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and analyzed by western blot with the indicated antibodies. Glyceraldehyde 3-phosphate dehydrogenase <t>(GAPDH)</t> was used as a loading control. The values of the band intensity represent the densitometric estimation of each band normalized to GAPDH.
    Horseradish Peroxidase Hrp Conjugated Gapdh Glyceraldehyde 3 Phosphate Dehydrogenase Monoclonal Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 79/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech mouse gapdh monoclonal
    Chronic constriction injury (CCI)-induced hyperalgesia and expression of <t>p53</t> and caspase-3 in dorsal root ganglion (DRG) neurons. (A) Compared to the sham group mice, the CCI mice had decreased paw withdrawal latency (PWL) from day 3 after surgery. (B) Western blot analysis of the expression of p53 and caspase-3 on day 7 after CCI or sham surgery. The fold change of p53 and caspase-3 was normalized to the glyceraldehyde 3-phosphate dehydrogenase <t>(GAPDH)</t> level. (C) Quantification of p53 and caspase-3 expression. Double immunofluorescence staining for p53 and caspase-3 in DRG neurons. (D) Representative immunofluorescence staining of p53 (red) and its colocalization with neurons (NeuN, green) in DRG tissues. (E) Representative immunofluorescence staining of caspase-3 (red) and its colocalization with neurons (NeuN, green) in DRG tissues. (F,G) Quantitative analysis of p53-positive and caspase-3-positive cells (%) in DRG tissues at 7 days after CCI. (H) Median fluorescence of NeuN. Magnification: 100× for all columns. Data are all expressed as mean ± SD. ∗ P
    Mouse Gapdh Monoclonal, supplied by Proteintech, used in various techniques. Bioz Stars score: 90/100, based on 1333 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore mouse anti glyceraldehyde 3 phosphate dehydrogenase
    Analysis of kazrin protein expression.  ( a ) Domain architecture of kazrinA and kazrinE. The N terminus of kazrinE is identical to kazrinA. ( b ) Purification of recombinant human kazrinA overexpressed in  Escherichia coli  as a glutathione- S -transferase (GST) fusion protein. ( c ) Lysates of normal human keratinocytes transfected with scrambled small interfering RNA (siControl) or pooled siRNAs specific for all isoforms of kazrin (siKazrinAll) were blotted with pan-kazrin antibody, rabbit pre-immune serum, or rabbit secondary antibody alone. ( d ) Pan-kazrin antibody crosslinked to protein G agarose beads was used to immunoprecipitate endogenous kazrin from lysates of normal human keratinocytes. ( e ,  f ) Pan-kazrin antibody detection of endogenous kazrin, kazrin β-galactosidase (β-gal) fusion protein, and the 28-kDa fragment encoded by kazrin exons 1–4 in lysates of wild-type (wt) mice or litter-matched gene trap (gt/gt) and conditional knockout (flx/flx) mice. simKazrinAll: wt/wt keratinocytes transfected with two pooled siRNAs (see also  Supplementary Figure S1c  online). Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a loading control for  c – f .
    Mouse Anti Glyceraldehyde 3 Phosphate Dehydrogenase, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 216 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech mouse anti human glyceraldehyde 3 phosphate dehydrogenase
    Hypoglycosylation of STT3B-dependent substrates in MagT1-depleted cells.  (A and C–F) HeLa cells were treated with NC or siRNAs specific for STT3A, STT3B, or MagT1 for 72 h. (A) HeLa cell extracts and cRM were resolved by PAGE in SDS and analyzed by protein immunoblotting using the specified antisera. MagT1-T7 expressed in HeLa cells verified recognition of MagT1 by the anti-MagT1 sera. Expression values relative to cells treated with the NC siRNA are for the displayed image, which is representative of two or more experiments. (B) Cell extracts prepared from STT3A-CDG, STT3B-CDG, and normal control (42F and 50F) fibroblasts were resolved by PAGE in SDS and analyzed by protein immunoblotting. The F0F1-ATPase α (A) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; B) served as gel loading controls. The asterisks in A and B designate a nonspecific product recognized by the anti-STT3B sera. Protein expression levels for OST subunits were normalized to the F 0 F 1 -ATPase α subunit loading control and are expressed relative to the NC siRNA lane. (C–F) After 48 h of siRNA treatment, cells were transfected with expression vectors for SHBG (C), or Hpx (Hpx-DDKHis or HpxΔ145-DDKHis; D) and pulse-chase labeled (4 min pulse, 20 min chase) after an additional 24 h. (E and F) After 72 h of siRNA treatment, cells were pulse labeled for 4 min and chased for 10 min. As indicated, samples were digested with EH after immunoprecipitation with anti-SHBG (C), anti-DDK (D), anti-SapD (E), or anti-granulin (F). Glycoforms resolved by PAGE in SDS are labeled to indicate the number of N-linked glycans. EH-digested proteins migrate slightly slower than the nonglycosylated protein because of the presence of a single residual GlcNAc residue at each site. Quantified values below gel lanes (C–F) are for the displayed image, which is representative of two or more experiments.
    Mouse Anti Human Glyceraldehyde 3 Phosphate Dehydrogenase, supplied by Proteintech, used in various techniques. Bioz Stars score: 78/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech horseradish hrp conjugated anti glyceraldehyde 3 phosphate dehydrogenase
    Hypoglycosylation of STT3B-dependent substrates in MagT1-depleted cells.  (A and C–F) HeLa cells were treated with NC or siRNAs specific for STT3A, STT3B, or MagT1 for 72 h. (A) HeLa cell extracts and cRM were resolved by PAGE in SDS and analyzed by protein immunoblotting using the specified antisera. MagT1-T7 expressed in HeLa cells verified recognition of MagT1 by the anti-MagT1 sera. Expression values relative to cells treated with the NC siRNA are for the displayed image, which is representative of two or more experiments. (B) Cell extracts prepared from STT3A-CDG, STT3B-CDG, and normal control (42F and 50F) fibroblasts were resolved by PAGE in SDS and analyzed by protein immunoblotting. The F0F1-ATPase α (A) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; B) served as gel loading controls. The asterisks in A and B designate a nonspecific product recognized by the anti-STT3B sera. Protein expression levels for OST subunits were normalized to the F 0 F 1 -ATPase α subunit loading control and are expressed relative to the NC siRNA lane. (C–F) After 48 h of siRNA treatment, cells were transfected with expression vectors for SHBG (C), or Hpx (Hpx-DDKHis or HpxΔ145-DDKHis; D) and pulse-chase labeled (4 min pulse, 20 min chase) after an additional 24 h. (E and F) After 72 h of siRNA treatment, cells were pulse labeled for 4 min and chased for 10 min. As indicated, samples were digested with EH after immunoprecipitation with anti-SHBG (C), anti-DDK (D), anti-SapD (E), or anti-granulin (F). Glycoforms resolved by PAGE in SDS are labeled to indicate the number of N-linked glycans. EH-digested proteins migrate slightly slower than the nonglycosylated protein because of the presence of a single residual GlcNAc residue at each site. Quantified values below gel lanes (C–F) are for the displayed image, which is representative of two or more experiments.
    Horseradish Hrp Conjugated Anti Glyceraldehyde 3 Phosphate Dehydrogenase, supplied by Proteintech, used in various techniques. Bioz Stars score: 77/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech anti gapdh
    Expression kinetics of endogenous interferon-inducible transmembrane protein 3 <t>(IFITM3)</t> protein modulated by virus infection. (A) A549, HeLa, and 293T cells were infected with VACV Tian Tan (VTT) at 0.1 PFU/cell for indicated time points after infection. Total cell proteins were extracted and the expression kinetics of IFITM3 and VTT D8 protein (*) were measured by Western blot and normalized to <t>GAPDH.</t> (B) Western blot analysis of IFITM1 and IFITM3 expression in HeLa cells treated with IFNα2b (10,000 U/mL) at indicated time point posttreatment. (C) Effect of IFNα2b at indicated concentrations on vaccinia virus infection in HeLa cells evaluated by flow cytometry and fluorescence microscope. (D,E) qPCR analysis of IFNα and IFNβ in HeLa and 293T cells infected by VTT infection at indicated time points postinfection. (F) Western blot analysis of IRF3 expression and phosphorylation in HeLa cells treated by VTT at indicated time points postinfection. ** P
    Anti Gapdh, supplied by Proteintech, used in various techniques. Bioz Stars score: 99/100, based on 1183 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech anti bax
    CXCR2 protects cells from undergoing stress-induced apoptosis. ( A ) Apoptosis in shNeg and shCXCR2 U2OS cells was measured 3 days after treatment with RSV (25 μM) by flow cytometry. ( B ) Cells were treated as in ( A ) and whole-cell extracts were collected for Western blot analysis using <t>BCL2</t> and <t>BAX</t> antibodies. ( C ) Downregulation of CXCR2 by siRNA as measured by RT-PCR and flow cytometry. U2OS cells were transfected with siRNA duplexes (200 nmol/L) specific to CXCR2 or scrabbled oligo in serum-free medium for 6 hours, then were incubated with complete medium for 24 h and then incubated with RSV for 3 days. ( D ) U2OS cells were treated the same as in ( C ) and apoptosis was measured by flow cytometry. ( E ) The U2OS cells were treated the same as in ( C ) and whole-cell extracts were collected for Western blot analysis using BCL2 antibodies. ( F ) Apoptosis in shNeg and shCXCR2 U2OS cells was measured by flow cytometry 2 days after treatment with H 2 O 2 (400 μM). Results shown are representative of three independent experiments. ( G ) The shNeg and shCXCR2 NHF cells were treated the same as in ( F ) and apoptosis was measured by flow cytometry. The numbers shown below Western blot images are means (first row) and SE (second row) of band intensities relative to control. Signals on the immunoblots were analyzed by ImageJ, normalized with that of GAPDH.
    Anti Bax, supplied by Proteintech, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech anti bax 505992 2 ig antibodies
    CXCR2 protects cells from undergoing stress-induced apoptosis. ( A ) Apoptosis in shNeg and shCXCR2 U2OS cells was measured 3 days after treatment with RSV (25 μM) by flow cytometry. ( B ) Cells were treated as in ( A ) and whole-cell extracts were collected for Western blot analysis using <t>BCL2</t> and <t>BAX</t> antibodies. ( C ) Downregulation of CXCR2 by siRNA as measured by RT-PCR and flow cytometry. U2OS cells were transfected with siRNA duplexes (200 nmol/L) specific to CXCR2 or scrabbled oligo in serum-free medium for 6 hours, then were incubated with complete medium for 24 h and then incubated with RSV for 3 days. ( D ) U2OS cells were treated the same as in ( C ) and apoptosis was measured by flow cytometry. ( E ) The U2OS cells were treated the same as in ( C ) and whole-cell extracts were collected for Western blot analysis using BCL2 antibodies. ( F ) Apoptosis in shNeg and shCXCR2 U2OS cells was measured by flow cytometry 2 days after treatment with H 2 O 2 (400 μM). Results shown are representative of three independent experiments. ( G ) The shNeg and shCXCR2 NHF cells were treated the same as in ( F ) and apoptosis was measured by flow cytometry. The numbers shown below Western blot images are means (first row) and SE (second row) of band intensities relative to control. Signals on the immunoblots were analyzed by ImageJ, normalized with that of GAPDH.
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    Proteintech rabbit anti bax
    Effects of aspirin on <t>Bcl-2,</t> <t>BAX,</t> PARP1, Cyclin D1 and P21 expression in RA-FLS. (A) After RA-FLS were exposed to various concentration of aspirin for 24 h, western blotting was used to determine Bcl-2, BAX, PARP1, Cyclin D1 and P21 protein levels. The corresponding internal control was GADPH. Changes in Bcl-2, BAX, PARP1, Cyclin D1 and P21 protein expression in RA-FLS treated with aspirin are shown: Bcl-2, PARP1, Cyclin D1 and P21 were decreased while BAX was increased, each with increasing aspirin concentration. (B) Bar graph shows the gray value analysis of Bcl-2, BAX, PARP1, Cyclin D1 and P21. Data are presented as the means ± SD (error bars) from three independent experiments. * P
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    Proteintech rabbit monoclonal anti bax
    Interactions of the autophagic and apoptotic effects of L02 cells induced by T-2 toxin. ( A ) Effect of RAPA on autophagy caused by T-2 toxin. L02 cells were pre-exposed to the autophagy stimulator RAPA (100 nM) for 24 h, and then co-treated with T-2 toxin (5 nM) for an additional 12 h. The levels of p62 and Beclin 1 proteins and the LC3-II/LC3-I ratio were analyzed by Western blotting (WB). ( B ) Effect of RAPA on apoptosis caused by T-2 toxin. L02 cells were pretreated with the autophagy stimulator RAPA (100 nM) for 24 h, and then co-treated with T-2 toxin (5 nM) for an additional 12 h. The levels of the apoptosis-related proteins of PARP-1 and caspase-3 as well as the <t>Bax/Bcl-2</t> ratio were analyzed by Western blotting (WB). ( C ) Effect of the autophagy inhibitor CQ on autophagy- and apoptosis-associated proteins in L02 cells exposed to T-2 toxin. L02 cells were pretreated with the autophagy inhibitor CQ (100 μM) for 1 h, and exposure to the inhibitor was continued during subsequent T-2 toxin (5 nM) treatment for 6 h. The LC3-II/LC3-I ratio, Beclin 1 level, and caspase-3 proteins were analyzed by Western blotting. ( D ) Effect of CQ on T-2 toxin-induced apoptosis. L02 cells were pretreated with the autophagy inhibitor CQ (100 μM) for 1 h, and then co-treated with T-2 toxin (5 nM) for an additional 6 h. Western blotting was used for the analysis of caspase-3 proteins. ( E ) The autophagic rate caused by T-2 toxin was greatly enhanced upon the inhibition of autophagy. The L02 cells were treated as described in ( D ) and cell apoptosis was detected using flow cytometry. Results are the mean ± SD, n = 3. ** represents p
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    Image Search Results


    Effects of DACT1 on β-catenin in myocardial cells. Effect of DACT1 overexpression on β-catenin concentration A) and distribution B) in H9C2 and HL-1 cells. C) Correlation between DACT1 and β-catenin expression in the myocardial tissues of patients with valvular heart disease. D) Colocation of DACT1 and β-catenin in the myocardial cells. H9C2 and HL-1 cells were overexpressed with the two isoforms of the DACT1 coding sequence, and then immunofluorescence was used to detect the DACT1 (DACT1-V1 or DACT1-V2) (OriGene) and β-catenin (CST) location. The secondary antibody Alexa Fluor 647 was used for DACT1 detection and Alexa Fluor 568 for β-catenin. The ZEN (Zeiss) software was used for further analysis. Alexa Fluor 647 was represented by the green color and Alexa Fluor 568 by the red color, while channel 488 was closed to avoid interference. DACT1=dishevelled binding antagonist of beta catenin 1; DAPI=2-(4-amidinophenyl)-6-indolecarbamidine dihydrochloride; GAPDH=glyceraldehyde-3-phosphate dehydrogenase

    Journal: Brazilian Journal of Cardiovascular Surgery

    Article Title: DACT1 Involvement in the Cytoskeletal Arrangement of Cardiomyocytes in Atrial Fibrillation by Regulating Cx43

    doi: 10.21470/1678-9741-2019-0033

    Figure Lengend Snippet: Effects of DACT1 on β-catenin in myocardial cells. Effect of DACT1 overexpression on β-catenin concentration A) and distribution B) in H9C2 and HL-1 cells. C) Correlation between DACT1 and β-catenin expression in the myocardial tissues of patients with valvular heart disease. D) Colocation of DACT1 and β-catenin in the myocardial cells. H9C2 and HL-1 cells were overexpressed with the two isoforms of the DACT1 coding sequence, and then immunofluorescence was used to detect the DACT1 (DACT1-V1 or DACT1-V2) (OriGene) and β-catenin (CST) location. The secondary antibody Alexa Fluor 647 was used for DACT1 detection and Alexa Fluor 568 for β-catenin. The ZEN (Zeiss) software was used for further analysis. Alexa Fluor 647 was represented by the green color and Alexa Fluor 568 by the red color, while channel 488 was closed to avoid interference. DACT1=dishevelled binding antagonist of beta catenin 1; DAPI=2-(4-amidinophenyl)-6-indolecarbamidine dihydrochloride; GAPDH=glyceraldehyde-3-phosphate dehydrogenase

    Article Snippet: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Proteintech) or β-actin (Wuhan GoodBio Tech.

    Techniques: Over Expression, Concentration Assay, Expressing, Sequencing, Immunofluorescence, Software, Binding Assay

    The effect of DACT1 on Cx43 in myocardial cells. The effect of DACT1 overexpression on Cx43 concentration A) and distribution B) in H9C2 and HL-1 cells. C) Cx43 expression in the myocardial tissues of patients with valvular heart disease. DACT1=dishevelled binding antagonist of beta catenin 1; GAPDH=glyceraldehyde-3-phosphate dehydrogenase

    Journal: Brazilian Journal of Cardiovascular Surgery

    Article Title: DACT1 Involvement in the Cytoskeletal Arrangement of Cardiomyocytes in Atrial Fibrillation by Regulating Cx43

    doi: 10.21470/1678-9741-2019-0033

    Figure Lengend Snippet: The effect of DACT1 on Cx43 in myocardial cells. The effect of DACT1 overexpression on Cx43 concentration A) and distribution B) in H9C2 and HL-1 cells. C) Cx43 expression in the myocardial tissues of patients with valvular heart disease. DACT1=dishevelled binding antagonist of beta catenin 1; GAPDH=glyceraldehyde-3-phosphate dehydrogenase

    Article Snippet: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Proteintech) or β-actin (Wuhan GoodBio Tech.

    Techniques: Over Expression, Concentration Assay, Expressing, Binding Assay

    Effect of lentivirus-mediated small hairpin RNA (shRNA) (S1) silencing on immature colon carcinoma transcript-1 (ICT1) expression in U251 cells. Notes: ( A ) Expression patterns of ICT1 in four glioblastoma multiforme (GBM) cell lines U251, U87, U373, and A172 determined by Western-blot analysis. ( B ) Green fluorescence protein (GFP) expression recorded under a fluorescence microscope. The representative pictures shown are from one of three independent experiments. ( C ) The messenger RNA levels of ICT1 in non-treated, constructed lentiviruses containing non-silencing small hairpin RNA (Lv-shCon)-treated and Lv-shICT1 (S1)-treated cells determined by RT-qPCR analysis. ( D ) The protein levels of ICT1 in non-treated, Lv-shCon-treated, and Lv-shICT1 (S1)-treated cells determined by Western-blot analysis. Scale bar in ( B ): 100 μm. Data are presented as mean ± standard deviation of three independent experiments performed in triplicate. *** P μ0.01 versus Lv-shCon. Abbreviations: con, control; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

    Journal: OncoTargets and therapy

    Article Title: Knockdown of immature colon carcinoma transcript-1 inhibits proliferation of glioblastoma multiforme cells through Gap 2/mitotic phase arrest

    doi: 10.2147/OTT.S75864

    Figure Lengend Snippet: Effect of lentivirus-mediated small hairpin RNA (shRNA) (S1) silencing on immature colon carcinoma transcript-1 (ICT1) expression in U251 cells. Notes: ( A ) Expression patterns of ICT1 in four glioblastoma multiforme (GBM) cell lines U251, U87, U373, and A172 determined by Western-blot analysis. ( B ) Green fluorescence protein (GFP) expression recorded under a fluorescence microscope. The representative pictures shown are from one of three independent experiments. ( C ) The messenger RNA levels of ICT1 in non-treated, constructed lentiviruses containing non-silencing small hairpin RNA (Lv-shCon)-treated and Lv-shICT1 (S1)-treated cells determined by RT-qPCR analysis. ( D ) The protein levels of ICT1 in non-treated, Lv-shCon-treated, and Lv-shICT1 (S1)-treated cells determined by Western-blot analysis. Scale bar in ( B ): 100 μm. Data are presented as mean ± standard deviation of three independent experiments performed in triplicate. *** P μ0.01 versus Lv-shCon. Abbreviations: con, control; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

    Article Snippet: The antibodies used were anti-ICT1 (1:1,000 dilution; Abgent, San Diego, CA, USA), anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:80,000 dilution; Proteintech Group, Inc, Chicago, IL, USA), and anti-rabbit HRP-IgG (1:5,000 dilution; Santa Cruz, Dallas, Texas, USA).

    Techniques: shRNA, Expressing, Western Blot, Fluorescence, Microscopy, Construct, Quantitative RT-PCR, Standard Deviation

    Effects of Lv-shICT1 (S2) on proliferation of U251 cells. The expression levels of immature colon carcinoma transcript-1 (ICT1) in non-treated, constructed lentiviruses containing non-silencing small hairpin RNA (Lv-shCon)-treated, and Lv-shICT1 (S2)-treated cells were determined by RT-qPCR analysis ( A ) and Western-blot analysis ( B ). ( C ) Statistical analysis of cell-proliferative rates in non-treated, Lv-shCon-treated, and Lv-shICT1 (S2)-treated cells. ( D ) Statistical analysis of colony numbers in non-treated, Lv-shCon-treated, and Lv-shICT1 (S2)-treated cells. Data are presented as mean ± SD of three independent experiments performed in triplicate. ** P μ0.01, *** P μ0.001 versus Lv-shCon. Abbreviations: con, control; OD, optical density; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

    Journal: OncoTargets and therapy

    Article Title: Knockdown of immature colon carcinoma transcript-1 inhibits proliferation of glioblastoma multiforme cells through Gap 2/mitotic phase arrest

    doi: 10.2147/OTT.S75864

    Figure Lengend Snippet: Effects of Lv-shICT1 (S2) on proliferation of U251 cells. The expression levels of immature colon carcinoma transcript-1 (ICT1) in non-treated, constructed lentiviruses containing non-silencing small hairpin RNA (Lv-shCon)-treated, and Lv-shICT1 (S2)-treated cells were determined by RT-qPCR analysis ( A ) and Western-blot analysis ( B ). ( C ) Statistical analysis of cell-proliferative rates in non-treated, Lv-shCon-treated, and Lv-shICT1 (S2)-treated cells. ( D ) Statistical analysis of colony numbers in non-treated, Lv-shCon-treated, and Lv-shICT1 (S2)-treated cells. Data are presented as mean ± SD of three independent experiments performed in triplicate. ** P μ0.01, *** P μ0.001 versus Lv-shCon. Abbreviations: con, control; OD, optical density; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

    Article Snippet: The antibodies used were anti-ICT1 (1:1,000 dilution; Abgent, San Diego, CA, USA), anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:80,000 dilution; Proteintech Group, Inc, Chicago, IL, USA), and anti-rabbit HRP-IgG (1:5,000 dilution; Santa Cruz, Dallas, Texas, USA).

    Techniques: Expressing, Construct, Quantitative RT-PCR, Western Blot

    The effect of silencing of the Slc7a6 gene on y + LAT2 protein level in astrocytes in relation to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein level. Upper panel shows representative Western blots. Results are mean ± SD; n = 4, * p

    Journal: International Journal of Molecular Sciences

    Article Title: Ammonia Reduces Intracellular Asymmetric Dimethylarginine in Cultured Astrocytes Stimulating Its y+LAT2 Carrier-Mediated Loss

    doi: 10.3390/ijms18112308

    Figure Lengend Snippet: The effect of silencing of the Slc7a6 gene on y + LAT2 protein level in astrocytes in relation to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein level. Upper panel shows representative Western blots. Results are mean ± SD; n = 4, * p

    Article Snippet: The first antibody was stripped off with 0.1 M glycine, pH 2.9, and second incubation was performed with an antibody against Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) 1 h incubation at 20–22 °C (1:7500, HRP-60004, ProteinTech, Manchester, UK).

    Techniques: Western Blot

    ISL induces cell cycle arrest in human endometrial cancer cells Cells were plated in 100 mm dishes at a density of 1 × 10 6 cells/plate in media supplemented with 10% FBS and allowed to adhere. The morphology of cells after treatment with vehicle or ISL (10, 25, 50 μΜ) in media containing 1% FBS for 48 h. ( A ) Ishikawa and ( B ) HEC-1A cells were stained with propidium iodide (PI), and cell cycle distribution was analyzed by flow cytometry. The vertical axis represents the number of cells, and the horizontal axis represents the intensity of PI staining. The cell cycle distribution is shown as a bar graph. The vertical numbers represent the cell population percentage in cell cycles S, G2, and G1phase, and the horizontal numbers represent the concentration of ISL. ( C ) Ishikawa and ( D ) HEC-1A cell lysates were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and analyzed by western blot with the indicated antibodies. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a loading control. The values of the band intensity represent the densitometric estimation of each band normalized to GAPDH.

    Journal: Oncotarget

    Article Title: Isoliquiritigenin induces apoptosis and autophagy and inhibits endometrial cancer growth in mice

    doi: 10.18632/oncotarget.12369

    Figure Lengend Snippet: ISL induces cell cycle arrest in human endometrial cancer cells Cells were plated in 100 mm dishes at a density of 1 × 10 6 cells/plate in media supplemented with 10% FBS and allowed to adhere. The morphology of cells after treatment with vehicle or ISL (10, 25, 50 μΜ) in media containing 1% FBS for 48 h. ( A ) Ishikawa and ( B ) HEC-1A cells were stained with propidium iodide (PI), and cell cycle distribution was analyzed by flow cytometry. The vertical axis represents the number of cells, and the horizontal axis represents the intensity of PI staining. The cell cycle distribution is shown as a bar graph. The vertical numbers represent the cell population percentage in cell cycles S, G2, and G1phase, and the horizontal numbers represent the concentration of ISL. ( C ) Ishikawa and ( D ) HEC-1A cell lysates were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and analyzed by western blot with the indicated antibodies. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a loading control. The values of the band intensity represent the densitometric estimation of each band normalized to GAPDH.

    Article Snippet: The following antibodies were used in this study: anti-light chain-3B (LC-3B) from Novus Biologicals (Littleton, CO, USA); anti-SQSTM1/p62 and anti-p21 Cip1 from GeneTex (Irvine, CA, USA); anti-caspase-3, anti-caspase-7, anti-cleaved-poly adenosine diphosphate-ribose polymerase (PARP), anti-p-extracellular signal regulated protein kinase (ERK)1/2, anti-ERK1/2, anti-p-γH2AX, anti-proliferating cell nuclear antigen (PCNA), and anti-p-p53 (Ser15) from Cell Signal Technology (Beverley, MA, USA); anti-horseradish peroxidase (HRP)-conjugated glyceraldehyde 3-phosphate dehydrogenase (GAPDH) from Proteintech (Rosemont, IL, USA); and goat anti-rabbit/mouse antibody IgG from Abcam (Cambridge, UK).

    Techniques: Staining, Flow Cytometry, Cytometry, Concentration Assay, Polyacrylamide Gel Electrophoresis, SDS Page, Western Blot

    Chronic constriction injury (CCI)-induced hyperalgesia and expression of p53 and caspase-3 in dorsal root ganglion (DRG) neurons. (A) Compared to the sham group mice, the CCI mice had decreased paw withdrawal latency (PWL) from day 3 after surgery. (B) Western blot analysis of the expression of p53 and caspase-3 on day 7 after CCI or sham surgery. The fold change of p53 and caspase-3 was normalized to the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) level. (C) Quantification of p53 and caspase-3 expression. Double immunofluorescence staining for p53 and caspase-3 in DRG neurons. (D) Representative immunofluorescence staining of p53 (red) and its colocalization with neurons (NeuN, green) in DRG tissues. (E) Representative immunofluorescence staining of caspase-3 (red) and its colocalization with neurons (NeuN, green) in DRG tissues. (F,G) Quantitative analysis of p53-positive and caspase-3-positive cells (%) in DRG tissues at 7 days after CCI. (H) Median fluorescence of NeuN. Magnification: 100× for all columns. Data are all expressed as mean ± SD. ∗ P

    Journal: Frontiers in Genetics

    Article Title: Bioinformatics Analysis Identifies p53 as a Candidate Prognostic Biomarker for Neuropathic Pain

    doi: 10.3389/fgene.2018.00320

    Figure Lengend Snippet: Chronic constriction injury (CCI)-induced hyperalgesia and expression of p53 and caspase-3 in dorsal root ganglion (DRG) neurons. (A) Compared to the sham group mice, the CCI mice had decreased paw withdrawal latency (PWL) from day 3 after surgery. (B) Western blot analysis of the expression of p53 and caspase-3 on day 7 after CCI or sham surgery. The fold change of p53 and caspase-3 was normalized to the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) level. (C) Quantification of p53 and caspase-3 expression. Double immunofluorescence staining for p53 and caspase-3 in DRG neurons. (D) Representative immunofluorescence staining of p53 (red) and its colocalization with neurons (NeuN, green) in DRG tissues. (E) Representative immunofluorescence staining of caspase-3 (red) and its colocalization with neurons (NeuN, green) in DRG tissues. (F,G) Quantitative analysis of p53-positive and caspase-3-positive cells (%) in DRG tissues at 7 days after CCI. (H) Median fluorescence of NeuN. Magnification: 100× for all columns. Data are all expressed as mean ± SD. ∗ P

    Article Snippet: The membranes were then incubated overnight at 4°C with rabbit anti-cleaved-caspase-3 primary antibody (ASP175; 1:1000; Cell Signaling Technology), mouse anti-p53 primary antibody (1C12; 1:1000; Cell Signaling Technology), or mouse anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody (60004-1-ig; 1:1000; Proteintech Group, Rosemont, IL, United States).

    Techniques: Expressing, Mouse Assay, Western Blot, Double Immunofluorescence Staining, Immunofluorescence, Staining, Fluorescence

    Inhibition of p53 by pifithrin-α (HY-15484) restrained the pain behaviors and expression of p53 and caspase-3 in dorsal root ganglion (DRG) neurons. (A) Thermal hyperalgesia after intrathecal injection of pifithrin-α (10 μg). Pifithrin-α attenuated chronic constriction injury (CCI)-induced thermal hyperalgesia, compared with that in the CCI + DMSO group. (B) Western blot analysis of the expression of p53 and caspase-3 after intrathecal administration of pifithrin-α on day 7 after CCI. The fold change of p53 and caspase-3 levels was normalized to the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) level. (C) Quantification of p53 and caspase-3 expression. Double immunofluorescence staining for p53 and caspase-3 in DRG neurons after intrathecal administration of pifithrin-α on day 7 after CCI. (D) Representative immunofluorescence staining of p53 (red) and its colocalization with neurons (NeuN, green) in DRG tissues. (E) Representative immunofluorescence staining of caspase-3 (red) and its colocalization with neurons (NeuN, green) in DRG tissues. (F,G) Quantitative analysis of p53-positive and caspase-3-positive cells (%) in DRG tissues at 7 days after intrathecal administration of pifithrin-α in the CCI model. (H) Median fluorescence of NeuN. Magnification: 100× for all columns. Data are all expressed as mean ± SD. # P

    Journal: Frontiers in Genetics

    Article Title: Bioinformatics Analysis Identifies p53 as a Candidate Prognostic Biomarker for Neuropathic Pain

    doi: 10.3389/fgene.2018.00320

    Figure Lengend Snippet: Inhibition of p53 by pifithrin-α (HY-15484) restrained the pain behaviors and expression of p53 and caspase-3 in dorsal root ganglion (DRG) neurons. (A) Thermal hyperalgesia after intrathecal injection of pifithrin-α (10 μg). Pifithrin-α attenuated chronic constriction injury (CCI)-induced thermal hyperalgesia, compared with that in the CCI + DMSO group. (B) Western blot analysis of the expression of p53 and caspase-3 after intrathecal administration of pifithrin-α on day 7 after CCI. The fold change of p53 and caspase-3 levels was normalized to the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) level. (C) Quantification of p53 and caspase-3 expression. Double immunofluorescence staining for p53 and caspase-3 in DRG neurons after intrathecal administration of pifithrin-α on day 7 after CCI. (D) Representative immunofluorescence staining of p53 (red) and its colocalization with neurons (NeuN, green) in DRG tissues. (E) Representative immunofluorescence staining of caspase-3 (red) and its colocalization with neurons (NeuN, green) in DRG tissues. (F,G) Quantitative analysis of p53-positive and caspase-3-positive cells (%) in DRG tissues at 7 days after intrathecal administration of pifithrin-α in the CCI model. (H) Median fluorescence of NeuN. Magnification: 100× for all columns. Data are all expressed as mean ± SD. # P

    Article Snippet: The membranes were then incubated overnight at 4°C with rabbit anti-cleaved-caspase-3 primary antibody (ASP175; 1:1000; Cell Signaling Technology), mouse anti-p53 primary antibody (1C12; 1:1000; Cell Signaling Technology), or mouse anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody (60004-1-ig; 1:1000; Proteintech Group, Rosemont, IL, United States).

    Techniques: Inhibition, Expressing, Injection, Western Blot, Double Immunofluorescence Staining, Immunofluorescence, Staining, Fluorescence

    Analysis of kazrin protein expression.  ( a ) Domain architecture of kazrinA and kazrinE. The N terminus of kazrinE is identical to kazrinA. ( b ) Purification of recombinant human kazrinA overexpressed in  Escherichia coli  as a glutathione- S -transferase (GST) fusion protein. ( c ) Lysates of normal human keratinocytes transfected with scrambled small interfering RNA (siControl) or pooled siRNAs specific for all isoforms of kazrin (siKazrinAll) were blotted with pan-kazrin antibody, rabbit pre-immune serum, or rabbit secondary antibody alone. ( d ) Pan-kazrin antibody crosslinked to protein G agarose beads was used to immunoprecipitate endogenous kazrin from lysates of normal human keratinocytes. ( e ,  f ) Pan-kazrin antibody detection of endogenous kazrin, kazrin β-galactosidase (β-gal) fusion protein, and the 28-kDa fragment encoded by kazrin exons 1–4 in lysates of wild-type (wt) mice or litter-matched gene trap (gt/gt) and conditional knockout (flx/flx) mice. simKazrinAll: wt/wt keratinocytes transfected with two pooled siRNAs (see also  Supplementary Figure S1c  online). Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a loading control for  c – f .

    Journal: The Journal of Investigative Dermatology

    Article Title: Exons 5-15 of Kazrin Are Dispensable for Murine Epidermal Morphogenesis and Homeostasis

    doi: 10.1038/jid.2012.110

    Figure Lengend Snippet: Analysis of kazrin protein expression. ( a ) Domain architecture of kazrinA and kazrinE. The N terminus of kazrinE is identical to kazrinA. ( b ) Purification of recombinant human kazrinA overexpressed in Escherichia coli as a glutathione- S -transferase (GST) fusion protein. ( c ) Lysates of normal human keratinocytes transfected with scrambled small interfering RNA (siControl) or pooled siRNAs specific for all isoforms of kazrin (siKazrinAll) were blotted with pan-kazrin antibody, rabbit pre-immune serum, or rabbit secondary antibody alone. ( d ) Pan-kazrin antibody crosslinked to protein G agarose beads was used to immunoprecipitate endogenous kazrin from lysates of normal human keratinocytes. ( e , f ) Pan-kazrin antibody detection of endogenous kazrin, kazrin β-galactosidase (β-gal) fusion protein, and the 28-kDa fragment encoded by kazrin exons 1–4 in lysates of wild-type (wt) mice or litter-matched gene trap (gt/gt) and conditional knockout (flx/flx) mice. simKazrinAll: wt/wt keratinocytes transfected with two pooled siRNAs (see also Supplementary Figure S1c online). Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a loading control for c – f .

    Article Snippet: The following antibodies were used (dilutions in brackets): rabbit anti-pan-kazrin (1:500), rabbit anti-peptide antibody to all kazrin isoforms (LS7, 1:500) , rabbit anti-kazrin (ProteinTech Group, Manchester, UK; 11572-1-AP; 1:500), rabbit pre-immune serum (1:500), mouse anti-α-tubulin (Sigma, Gillingham, Dorset, UK; T6199; 1:2000), and mouse-anti-glyceraldehyde 3-phosphate dehydrogenase (Millipore, Watford, UK; MAB374; 1:500).

    Techniques: Expressing, Purification, Recombinant, Transfection, Small Interfering RNA, Mouse Assay, Knock-Out

    Generation of kazrin β-galactosidase (β-gal) gene-trap (gt/gt) mouse and conditional knockout (flx/flx) mice.  ( a ) Exon structure of mouse kazrin. Blue box represents insertion of the neomycin resistance gene ( β-geo ) cassette in the kazrin β-gal gt/gt mouse. Red triangles indicate loxP sites for removing exon 5 in the kazrin flx/flx mice. ( b ) Predicted domain architecture of kazrin-β-geo fusion protein in the kazrin β-gal (top) gt/gt mouse and kazrin fragment expressed in the (bottom) kazrin flx/flx mouse. ( c ) RT-PCR amplification of transcripts upstream (exons 2–3) or downstream (exons 4–5) of the gene trap. RNA in each lane is from a single representative litter-matched mouse of the genotype indicated. ( d ) Quantitative RT-PCR of transcripts upstream (exons 2–3) and downstream (exons 6–7) of the gene trap. Top: Expression values relative to glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Each bar shows the mean and standard deviation of at least three mice per genotype. Bottom: Ratio (%) of upstream and downstream transcripts from the top panel. Primer positions upstream and downstream of the  β-geo  cassette are indicated by the orange and green brackets, respectively, in  a . ( e ) Alignment of predicted amino-acid sequences encoded by exons 1 and 2 of human kazrinA and mouse kazrin. ( f ) RT-PCR amplification of exons 1–4, 1–5, 2–4, or 2–5 of mouse kazrinA. Positive (GAPDH) and negative (water) controls are shown.  β-geo , β-galactosidase fused to a neomycin resistance gene; Ex, exon.

    Journal: The Journal of Investigative Dermatology

    Article Title: Exons 5-15 of Kazrin Are Dispensable for Murine Epidermal Morphogenesis and Homeostasis

    doi: 10.1038/jid.2012.110

    Figure Lengend Snippet: Generation of kazrin β-galactosidase (β-gal) gene-trap (gt/gt) mouse and conditional knockout (flx/flx) mice. ( a ) Exon structure of mouse kazrin. Blue box represents insertion of the neomycin resistance gene ( β-geo ) cassette in the kazrin β-gal gt/gt mouse. Red triangles indicate loxP sites for removing exon 5 in the kazrin flx/flx mice. ( b ) Predicted domain architecture of kazrin-β-geo fusion protein in the kazrin β-gal (top) gt/gt mouse and kazrin fragment expressed in the (bottom) kazrin flx/flx mouse. ( c ) RT-PCR amplification of transcripts upstream (exons 2–3) or downstream (exons 4–5) of the gene trap. RNA in each lane is from a single representative litter-matched mouse of the genotype indicated. ( d ) Quantitative RT-PCR of transcripts upstream (exons 2–3) and downstream (exons 6–7) of the gene trap. Top: Expression values relative to glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Each bar shows the mean and standard deviation of at least three mice per genotype. Bottom: Ratio (%) of upstream and downstream transcripts from the top panel. Primer positions upstream and downstream of the β-geo cassette are indicated by the orange and green brackets, respectively, in a . ( e ) Alignment of predicted amino-acid sequences encoded by exons 1 and 2 of human kazrinA and mouse kazrin. ( f ) RT-PCR amplification of exons 1–4, 1–5, 2–4, or 2–5 of mouse kazrinA. Positive (GAPDH) and negative (water) controls are shown. β-geo , β-galactosidase fused to a neomycin resistance gene; Ex, exon.

    Article Snippet: The following antibodies were used (dilutions in brackets): rabbit anti-pan-kazrin (1:500), rabbit anti-peptide antibody to all kazrin isoforms (LS7, 1:500) , rabbit anti-kazrin (ProteinTech Group, Manchester, UK; 11572-1-AP; 1:500), rabbit pre-immune serum (1:500), mouse anti-α-tubulin (Sigma, Gillingham, Dorset, UK; T6199; 1:2000), and mouse-anti-glyceraldehyde 3-phosphate dehydrogenase (Millipore, Watford, UK; MAB374; 1:500).

    Techniques: Knock-Out, Mouse Assay, Reverse Transcription Polymerase Chain Reaction, Amplification, Quantitative RT-PCR, Expressing, Standard Deviation

    Hypoglycosylation of STT3B-dependent substrates in MagT1-depleted cells.  (A and C–F) HeLa cells were treated with NC or siRNAs specific for STT3A, STT3B, or MagT1 for 72 h. (A) HeLa cell extracts and cRM were resolved by PAGE in SDS and analyzed by protein immunoblotting using the specified antisera. MagT1-T7 expressed in HeLa cells verified recognition of MagT1 by the anti-MagT1 sera. Expression values relative to cells treated with the NC siRNA are for the displayed image, which is representative of two or more experiments. (B) Cell extracts prepared from STT3A-CDG, STT3B-CDG, and normal control (42F and 50F) fibroblasts were resolved by PAGE in SDS and analyzed by protein immunoblotting. The F0F1-ATPase α (A) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; B) served as gel loading controls. The asterisks in A and B designate a nonspecific product recognized by the anti-STT3B sera. Protein expression levels for OST subunits were normalized to the F 0 F 1 -ATPase α subunit loading control and are expressed relative to the NC siRNA lane. (C–F) After 48 h of siRNA treatment, cells were transfected with expression vectors for SHBG (C), or Hpx (Hpx-DDKHis or HpxΔ145-DDKHis; D) and pulse-chase labeled (4 min pulse, 20 min chase) after an additional 24 h. (E and F) After 72 h of siRNA treatment, cells were pulse labeled for 4 min and chased for 10 min. As indicated, samples were digested with EH after immunoprecipitation with anti-SHBG (C), anti-DDK (D), anti-SapD (E), or anti-granulin (F). Glycoforms resolved by PAGE in SDS are labeled to indicate the number of N-linked glycans. EH-digested proteins migrate slightly slower than the nonglycosylated protein because of the presence of a single residual GlcNAc residue at each site. Quantified values below gel lanes (C–F) are for the displayed image, which is representative of two or more experiments.

    Journal: The Journal of Cell Biology

    Article Title: Oxidoreductase activity is necessary for N-glycosylation of cysteine-proximal acceptor sites in glycoproteins

    doi: 10.1083/jcb.201404083

    Figure Lengend Snippet: Hypoglycosylation of STT3B-dependent substrates in MagT1-depleted cells. (A and C–F) HeLa cells were treated with NC or siRNAs specific for STT3A, STT3B, or MagT1 for 72 h. (A) HeLa cell extracts and cRM were resolved by PAGE in SDS and analyzed by protein immunoblotting using the specified antisera. MagT1-T7 expressed in HeLa cells verified recognition of MagT1 by the anti-MagT1 sera. Expression values relative to cells treated with the NC siRNA are for the displayed image, which is representative of two or more experiments. (B) Cell extracts prepared from STT3A-CDG, STT3B-CDG, and normal control (42F and 50F) fibroblasts were resolved by PAGE in SDS and analyzed by protein immunoblotting. The F0F1-ATPase α (A) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; B) served as gel loading controls. The asterisks in A and B designate a nonspecific product recognized by the anti-STT3B sera. Protein expression levels for OST subunits were normalized to the F 0 F 1 -ATPase α subunit loading control and are expressed relative to the NC siRNA lane. (C–F) After 48 h of siRNA treatment, cells were transfected with expression vectors for SHBG (C), or Hpx (Hpx-DDKHis or HpxΔ145-DDKHis; D) and pulse-chase labeled (4 min pulse, 20 min chase) after an additional 24 h. (E and F) After 72 h of siRNA treatment, cells were pulse labeled for 4 min and chased for 10 min. As indicated, samples were digested with EH after immunoprecipitation with anti-SHBG (C), anti-DDK (D), anti-SapD (E), or anti-granulin (F). Glycoforms resolved by PAGE in SDS are labeled to indicate the number of N-linked glycans. EH-digested proteins migrate slightly slower than the nonglycosylated protein because of the presence of a single residual GlcNAc residue at each site. Quantified values below gel lanes (C–F) are for the displayed image, which is representative of two or more experiments.

    Article Snippet: Rabbit anti–human MagT1 (17430), rabbit anti–human TUSC3 (16039), and mouse anti–human glyceraldehyde 3-phosphate dehydrogenase (GAPDH; 60004) were obtained from Proteintech Group, Inc.

    Techniques: Polyacrylamide Gel Electrophoresis, Expressing, Transfection, Pulse Chase, Labeling, Immunoprecipitation

    Expression kinetics of endogenous interferon-inducible transmembrane protein 3 (IFITM3) protein modulated by virus infection. (A) A549, HeLa, and 293T cells were infected with VACV Tian Tan (VTT) at 0.1 PFU/cell for indicated time points after infection. Total cell proteins were extracted and the expression kinetics of IFITM3 and VTT D8 protein (*) were measured by Western blot and normalized to GAPDH. (B) Western blot analysis of IFITM1 and IFITM3 expression in HeLa cells treated with IFNα2b (10,000 U/mL) at indicated time point posttreatment. (C) Effect of IFNα2b at indicated concentrations on vaccinia virus infection in HeLa cells evaluated by flow cytometry and fluorescence microscope. (D,E) qPCR analysis of IFNα and IFNβ in HeLa and 293T cells infected by VTT infection at indicated time points postinfection. (F) Western blot analysis of IRF3 expression and phosphorylation in HeLa cells treated by VTT at indicated time points postinfection. ** P

    Journal: Frontiers in Immunology

    Article Title: The Host Restriction Factor Interferon-Inducible Transmembrane Protein 3 Inhibits Vaccinia Virus Infection

    doi: 10.3389/fimmu.2018.00228

    Figure Lengend Snippet: Expression kinetics of endogenous interferon-inducible transmembrane protein 3 (IFITM3) protein modulated by virus infection. (A) A549, HeLa, and 293T cells were infected with VACV Tian Tan (VTT) at 0.1 PFU/cell for indicated time points after infection. Total cell proteins were extracted and the expression kinetics of IFITM3 and VTT D8 protein (*) were measured by Western blot and normalized to GAPDH. (B) Western blot analysis of IFITM1 and IFITM3 expression in HeLa cells treated with IFNα2b (10,000 U/mL) at indicated time point posttreatment. (C) Effect of IFNα2b at indicated concentrations on vaccinia virus infection in HeLa cells evaluated by flow cytometry and fluorescence microscope. (D,E) qPCR analysis of IFNα and IFNβ in HeLa and 293T cells infected by VTT infection at indicated time points postinfection. (F) Western blot analysis of IRF3 expression and phosphorylation in HeLa cells treated by VTT at indicated time points postinfection. ** P

    Article Snippet: Anti-FLAG M2 antibody was purchased from Sigma-Aldrich; anti-IFITM3, anti-GAPDH (glyceraldehyde 3-phosphate dehydrogenase), and anti-β-actin antibodies from Proteintech (Chicago, IL, USA); Anti-VACV D8 (WR113) antibody was purchased from Immune Tech (New York, NY, USA).

    Techniques: Expressing, Infection, Western Blot, Flow Cytometry, Cytometry, Fluorescence, Microscopy, Real-time Polymerase Chain Reaction

    CXCR2 protects cells from undergoing stress-induced apoptosis. ( A ) Apoptosis in shNeg and shCXCR2 U2OS cells was measured 3 days after treatment with RSV (25 μM) by flow cytometry. ( B ) Cells were treated as in ( A ) and whole-cell extracts were collected for Western blot analysis using BCL2 and BAX antibodies. ( C ) Downregulation of CXCR2 by siRNA as measured by RT-PCR and flow cytometry. U2OS cells were transfected with siRNA duplexes (200 nmol/L) specific to CXCR2 or scrabbled oligo in serum-free medium for 6 hours, then were incubated with complete medium for 24 h and then incubated with RSV for 3 days. ( D ) U2OS cells were treated the same as in ( C ) and apoptosis was measured by flow cytometry. ( E ) The U2OS cells were treated the same as in ( C ) and whole-cell extracts were collected for Western blot analysis using BCL2 antibodies. ( F ) Apoptosis in shNeg and shCXCR2 U2OS cells was measured by flow cytometry 2 days after treatment with H 2 O 2 (400 μM). Results shown are representative of three independent experiments. ( G ) The shNeg and shCXCR2 NHF cells were treated the same as in ( F ) and apoptosis was measured by flow cytometry. The numbers shown below Western blot images are means (first row) and SE (second row) of band intensities relative to control. Signals on the immunoblots were analyzed by ImageJ, normalized with that of GAPDH.

    Journal: Scientific Reports

    Article Title: Resveratrol sequentially induces replication and oxidative stresses to drive p53-CXCR2 mediated cellular senescence in cancer cells

    doi: 10.1038/s41598-017-00315-4

    Figure Lengend Snippet: CXCR2 protects cells from undergoing stress-induced apoptosis. ( A ) Apoptosis in shNeg and shCXCR2 U2OS cells was measured 3 days after treatment with RSV (25 μM) by flow cytometry. ( B ) Cells were treated as in ( A ) and whole-cell extracts were collected for Western blot analysis using BCL2 and BAX antibodies. ( C ) Downregulation of CXCR2 by siRNA as measured by RT-PCR and flow cytometry. U2OS cells were transfected with siRNA duplexes (200 nmol/L) specific to CXCR2 or scrabbled oligo in serum-free medium for 6 hours, then were incubated with complete medium for 24 h and then incubated with RSV for 3 days. ( D ) U2OS cells were treated the same as in ( C ) and apoptosis was measured by flow cytometry. ( E ) The U2OS cells were treated the same as in ( C ) and whole-cell extracts were collected for Western blot analysis using BCL2 antibodies. ( F ) Apoptosis in shNeg and shCXCR2 U2OS cells was measured by flow cytometry 2 days after treatment with H 2 O 2 (400 μM). Results shown are representative of three independent experiments. ( G ) The shNeg and shCXCR2 NHF cells were treated the same as in ( F ) and apoptosis was measured by flow cytometry. The numbers shown below Western blot images are means (first row) and SE (second row) of band intensities relative to control. Signals on the immunoblots were analyzed by ImageJ, normalized with that of GAPDH.

    Article Snippet: The antibodies used for Western blot were anti-p-p53 (No. 9284s, Cell Signaling; 1:1000), anti-p53 (sc-126, Santa Cruz; 1:1000), anti-p-CHK1 (ab58567, abcam; 1:1000), anti-CHK1 (No.2360, Cell Signaling Technology; 1:1000),anti-p-ATM (No. 5883s, Cell Signaling Technology; 1:1000), anti-BCL2 (60178-1-Ig, proteintech; 1:1000), anti-BAX (60267-1-lg,proteintech; 1:1000), anti-GAPDH (Chemicon; 1:10000).

    Techniques: Flow Cytometry, Cytometry, Western Blot, Reverse Transcription Polymerase Chain Reaction, Transfection, Incubation

    Bcl-2 overexpression inhibits the activation of the mitochondrial apoptosis pathway by cisplatin in SKOV3 cells. (A and B) Pc-SKOV3 and Bcl-2-SKOV3 cells were treated with 6 µg/ml cisplatin for 6 h and then stained with MitoPotential dye and 7-AAD to assess the Δψm by flow cytometry. (C and D) Western blot analysis of cytochrome c , Bcl-2, Bax, cleaved caspase-9 and cleaved caspase-3 levels in pc-SKOV3 and Bcl-2-SKOV3 cells after cisplatin treatment. The results are the mean ± SD of three independent experiments. *P

    Journal: Oncology Reports

    Article Title: Bcl-2 overexpression reduces cisplatin cytotoxicity by decreasing ER-mitochondrial Ca2+ signaling in SKOV3 cells

    doi: 10.3892/or.2017.6164

    Figure Lengend Snippet: Bcl-2 overexpression inhibits the activation of the mitochondrial apoptosis pathway by cisplatin in SKOV3 cells. (A and B) Pc-SKOV3 and Bcl-2-SKOV3 cells were treated with 6 µg/ml cisplatin for 6 h and then stained with MitoPotential dye and 7-AAD to assess the Δψm by flow cytometry. (C and D) Western blot analysis of cytochrome c , Bcl-2, Bax, cleaved caspase-9 and cleaved caspase-3 levels in pc-SKOV3 and Bcl-2-SKOV3 cells after cisplatin treatment. The results are the mean ± SD of three independent experiments. *P

    Article Snippet: Anti-β-actin (60008–1-Ig; murine Ab; 1:2,000 dilution), anti-Bax (50599–2-Ig; rabbit Ab; 1:2,000 dilution), anti-Bcl-2 (12789–1-AP; rabbit Ab; 1:1,000 dilution), anti-cytochrome c (cyto c ) (10993–1-AP; rabbit Ab; 1:1,000 dilution), anti-Grp78/BIP (11587–1-AP; rabbit Ab; 1:1,000 dilution), peroxidase-conjugated AffiniPure goat anti-mouse IgG (H+L) (SA00001-1; goat Ab; 1:2,000 dilution) and peroxidase-conjugated AffiniPure goat anti-rabbit IgG (H+L) (SA00001-2; goat Ab; 1:2,000 dilution) Abs were purchased from ProteinTech Group, Inc. (Chicago, IL, USA).

    Techniques: Over Expression, Activation Assay, Staining, Flow Cytometry, Cytometry, Western Blot

    Antiapoptosis effects of methane-rich saline on septic AKI. (a) TUNEL assay was performed on kidney tissue slices. (b) The quantity of TUNEL-positive cells was counted in a high-power field. (c) The kidney protein expression levels of Bcl-2 and Bax were detected by Western blot, and the relative band intensities (fold of the sham group) were shown in (d). (e) The protein levels of cleaved caspase-3 and cleaved PARP were detected, and the relative band intensities (fold of the sham group) were shown in (f). ∗ P

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Methane-Rich Saline Ameliorates Sepsis-Induced Acute Kidney Injury through Anti-Inflammation, Antioxidative, and Antiapoptosis Effects by Regulating Endoplasmic Reticulum Stress

    doi: 10.1155/2018/4756846

    Figure Lengend Snippet: Antiapoptosis effects of methane-rich saline on septic AKI. (a) TUNEL assay was performed on kidney tissue slices. (b) The quantity of TUNEL-positive cells was counted in a high-power field. (c) The kidney protein expression levels of Bcl-2 and Bax were detected by Western blot, and the relative band intensities (fold of the sham group) were shown in (d). (e) The protein levels of cleaved caspase-3 and cleaved PARP were detected, and the relative band intensities (fold of the sham group) were shown in (f). ∗ P

    Article Snippet: The resulting blots were blocked with 8% skim milk and incubated with an anti-GRP78 antibody (1 : 5000; Proteintech, China), an anti-ATF4 antibody (1 : 1000; Proteintech, China), anti-caspase-12 antibodies (1 : 1000; Cell Signaling Technology (CST), USA), anti-IL-1β antibodies (1 : 1000; Abcam, USA), anti-TNF-α (1 : 500; Abcam, USA), anti-Bcl2 antibodies (1 : 1000; Biosis, China), anti-Bax antibodies (1 : 2000; Proteintech, China), anti-cleaved-caspase-3 antibodies (1 : 1000; Cell Signaling Technology (CST), USA), anti-PARP antibodies (1 : 1000; Cell Signaling Technology (CST), USA), and anti-β -actin antibodies (1 : 10000; Santa Cruz Biotechnology, USA) overnight at 4°C.

    Techniques: TUNEL Assay, Expressing, Western Blot

    Effects of aspirin on Bcl-2, BAX, PARP1, Cyclin D1 and P21 expression in RA-FLS. (A) After RA-FLS were exposed to various concentration of aspirin for 24 h, western blotting was used to determine Bcl-2, BAX, PARP1, Cyclin D1 and P21 protein levels. The corresponding internal control was GADPH. Changes in Bcl-2, BAX, PARP1, Cyclin D1 and P21 protein expression in RA-FLS treated with aspirin are shown: Bcl-2, PARP1, Cyclin D1 and P21 were decreased while BAX was increased, each with increasing aspirin concentration. (B) Bar graph shows the gray value analysis of Bcl-2, BAX, PARP1, Cyclin D1 and P21. Data are presented as the means ± SD (error bars) from three independent experiments. * P

    Journal: International Journal of Molecular Medicine

    Article Title: Aspirin promotes apoptosis and inhibits proliferation by blocking G0/G1 into S phase in rheumatoid arthritis fibroblast-like synoviocytes via downregulation of JAK/STAT3 and NF-κB signaling pathway

    doi: 10.3892/ijmm.2018.3883

    Figure Lengend Snippet: Effects of aspirin on Bcl-2, BAX, PARP1, Cyclin D1 and P21 expression in RA-FLS. (A) After RA-FLS were exposed to various concentration of aspirin for 24 h, western blotting was used to determine Bcl-2, BAX, PARP1, Cyclin D1 and P21 protein levels. The corresponding internal control was GADPH. Changes in Bcl-2, BAX, PARP1, Cyclin D1 and P21 protein expression in RA-FLS treated with aspirin are shown: Bcl-2, PARP1, Cyclin D1 and P21 were decreased while BAX was increased, each with increasing aspirin concentration. (B) Bar graph shows the gray value analysis of Bcl-2, BAX, PARP1, Cyclin D1 and P21. Data are presented as the means ± SD (error bars) from three independent experiments. * P

    Article Snippet: The membranes were blocked with 5% BSA in TBS-T for 1 h, and then incubated with rabbit anti-Bax (1:2,000 dilution, Proteintech Group; #S0599-2-lg), anti-Bcl-2 (1:1,000 dilution, Santa Cruz Biotechnology, Inc.; #sc-7382), anti-PARP1 (1:2,000 dilution, Proteintech Group; #13371-1-AP), anti-P21 (1:1,000 dilution, #EPR3993), anti-cyclin D1 (1:10,000 dilution, #EPR2241; both from Abcam), anti-P65 (1:2,000 dilution; #8242), anti-p-P65 (1:2,000 dilution; #3033; both from Cell Signaling Technology, Inc.), anti-P50/P105 (1:1,000 dilution, Abcam: #E381), anti-p-P50/105 (1:1,000 dilution, Affinity Company: #AF3219), anti-STAT3 (1:2,000 dilution, #EPR787Y) and anti-p-STAT3 (1:200,000 dilution, #EP2147Y; both from Abcam) antibodies for 2 h at room temperature.

    Techniques: Expressing, Concentration Assay, Western Blot

    Interactions of the autophagic and apoptotic effects of L02 cells induced by T-2 toxin. ( A ) Effect of RAPA on autophagy caused by T-2 toxin. L02 cells were pre-exposed to the autophagy stimulator RAPA (100 nM) for 24 h, and then co-treated with T-2 toxin (5 nM) for an additional 12 h. The levels of p62 and Beclin 1 proteins and the LC3-II/LC3-I ratio were analyzed by Western blotting (WB). ( B ) Effect of RAPA on apoptosis caused by T-2 toxin. L02 cells were pretreated with the autophagy stimulator RAPA (100 nM) for 24 h, and then co-treated with T-2 toxin (5 nM) for an additional 12 h. The levels of the apoptosis-related proteins of PARP-1 and caspase-3 as well as the Bax/Bcl-2 ratio were analyzed by Western blotting (WB). ( C ) Effect of the autophagy inhibitor CQ on autophagy- and apoptosis-associated proteins in L02 cells exposed to T-2 toxin. L02 cells were pretreated with the autophagy inhibitor CQ (100 μM) for 1 h, and exposure to the inhibitor was continued during subsequent T-2 toxin (5 nM) treatment for 6 h. The LC3-II/LC3-I ratio, Beclin 1 level, and caspase-3 proteins were analyzed by Western blotting. ( D ) Effect of CQ on T-2 toxin-induced apoptosis. L02 cells were pretreated with the autophagy inhibitor CQ (100 μM) for 1 h, and then co-treated with T-2 toxin (5 nM) for an additional 6 h. Western blotting was used for the analysis of caspase-3 proteins. ( E ) The autophagic rate caused by T-2 toxin was greatly enhanced upon the inhibition of autophagy. The L02 cells were treated as described in ( D ) and cell apoptosis was detected using flow cytometry. Results are the mean ± SD, n = 3. ** represents p

    Journal: Toxins

    Article Title: Autophagy and Apoptosis Interact to Modulate T-2 Toxin-Induced Toxicity in Liver Cells

    doi: 10.3390/toxins11010045

    Figure Lengend Snippet: Interactions of the autophagic and apoptotic effects of L02 cells induced by T-2 toxin. ( A ) Effect of RAPA on autophagy caused by T-2 toxin. L02 cells were pre-exposed to the autophagy stimulator RAPA (100 nM) for 24 h, and then co-treated with T-2 toxin (5 nM) for an additional 12 h. The levels of p62 and Beclin 1 proteins and the LC3-II/LC3-I ratio were analyzed by Western blotting (WB). ( B ) Effect of RAPA on apoptosis caused by T-2 toxin. L02 cells were pretreated with the autophagy stimulator RAPA (100 nM) for 24 h, and then co-treated with T-2 toxin (5 nM) for an additional 12 h. The levels of the apoptosis-related proteins of PARP-1 and caspase-3 as well as the Bax/Bcl-2 ratio were analyzed by Western blotting (WB). ( C ) Effect of the autophagy inhibitor CQ on autophagy- and apoptosis-associated proteins in L02 cells exposed to T-2 toxin. L02 cells were pretreated with the autophagy inhibitor CQ (100 μM) for 1 h, and exposure to the inhibitor was continued during subsequent T-2 toxin (5 nM) treatment for 6 h. The LC3-II/LC3-I ratio, Beclin 1 level, and caspase-3 proteins were analyzed by Western blotting. ( D ) Effect of CQ on T-2 toxin-induced apoptosis. L02 cells were pretreated with the autophagy inhibitor CQ (100 μM) for 1 h, and then co-treated with T-2 toxin (5 nM) for an additional 6 h. Western blotting was used for the analysis of caspase-3 proteins. ( E ) The autophagic rate caused by T-2 toxin was greatly enhanced upon the inhibition of autophagy. The L02 cells were treated as described in ( D ) and cell apoptosis was detected using flow cytometry. Results are the mean ± SD, n = 3. ** represents p

    Article Snippet: Antibodies of mouse monoclonal anti-β-actin, rabbit monoclonal anti-Bcl-2, rabbit monoclonal anti-Bax, rabbit monoclonal anti-PARP-1, rabbit monoclonal anti-caspase-3, rabbit monoclonal anti-P62, rabbit monoclonal anti-Beclin 1, rabbit monoclonal anti-LC3, goat anti-rabbit IgG-HRP, and goat anti-mouse IgG-HRP were purchased from Proteintech (Rosemont, IL, USA).

    Techniques: Western Blot, Inhibition, Flow Cytometry, Cytometry

    Apoptotic effects of L02 cells caused by T-2 toxin. ( A ) Increased dose-dependent apoptosis caused by T-2 toxin. The L02 cells were exposed to 0, 0.2, 1, and 5 nM T-2 toxin for 12 h, and the poly(ADP-ribose) polymerase 1 (PARP-1) and caspase-3 cleavage as well as the Bax/Bcl-2 ratio were detected using Western blotting (WB). Results are the mean ± SD, n = 3. ** represents p

    Journal: Toxins

    Article Title: Autophagy and Apoptosis Interact to Modulate T-2 Toxin-Induced Toxicity in Liver Cells

    doi: 10.3390/toxins11010045

    Figure Lengend Snippet: Apoptotic effects of L02 cells caused by T-2 toxin. ( A ) Increased dose-dependent apoptosis caused by T-2 toxin. The L02 cells were exposed to 0, 0.2, 1, and 5 nM T-2 toxin for 12 h, and the poly(ADP-ribose) polymerase 1 (PARP-1) and caspase-3 cleavage as well as the Bax/Bcl-2 ratio were detected using Western blotting (WB). Results are the mean ± SD, n = 3. ** represents p

    Article Snippet: Antibodies of mouse monoclonal anti-β-actin, rabbit monoclonal anti-Bcl-2, rabbit monoclonal anti-Bax, rabbit monoclonal anti-PARP-1, rabbit monoclonal anti-caspase-3, rabbit monoclonal anti-P62, rabbit monoclonal anti-Beclin 1, rabbit monoclonal anti-LC3, goat anti-rabbit IgG-HRP, and goat anti-mouse IgG-HRP were purchased from Proteintech (Rosemont, IL, USA).

    Techniques: Western Blot