rabbit anti human ddit3 Search Results


c ebp homologous protein chop  (Bioss)
94
Bioss c ebp homologous protein chop
Expression of CREB3L1, GRP78 and <t>CHOP</t> in World Health Organization grade I-Ⅳ glioma tissues detected by immunohistochemistry (magnification, x400). Values are expressed as the mean ± standard deviation (n=3). **P<0.01 vs. grade I. CREB3L1, cyclic adenosine monophosphate responsive element binding protein 3 like 1; CHOP, <t>C/EBP-homologous</t> protein; GRP78, glucose-regulated protein, 78 kDa.
C Ebp Homologous Protein Chop, supplied by Bioss, 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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gene exp ddit3 mm00492097 m1  (Thermo Fisher)
95
Thermo Fisher gene exp ddit3 mm00492097 m1
Expression of CREB3L1, GRP78 and <t>CHOP</t> in World Health Organization grade I-Ⅳ glioma tissues detected by immunohistochemistry (magnification, x400). Values are expressed as the mean ± standard deviation (n=3). **P<0.01 vs. grade I. CREB3L1, cyclic adenosine monophosphate responsive element binding protein 3 like 1; CHOP, <t>C/EBP-homologous</t> protein; GRP78, glucose-regulated protein, 78 kDa.
Gene Exp Ddit3 Mm00492097 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 95 stars, based on 1 article reviews
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anti ((chop, affinity, af5280)  (Proteintech)
96
Proteintech anti ((chop, affinity, af5280)
Expression of CREB3L1, GRP78 and <t>CHOP</t> in World Health Organization grade I-Ⅳ glioma tissues detected by immunohistochemistry (magnification, x400). Values are expressed as the mean ± standard deviation (n=3). **P<0.01 vs. grade I. CREB3L1, cyclic adenosine monophosphate responsive element binding protein 3 like 1; CHOP, <t>C/EBP-homologous</t> protein; GRP78, glucose-regulated protein, 78 kDa.
Anti ((Chop, Affinity, Af5280), 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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chop  (Proteintech)
96
Proteintech chop
Expression of CREB3L1, GRP78 and <t>CHOP</t> in World Health Organization grade I-Ⅳ glioma tissues detected by immunohistochemistry (magnification, x400). Values are expressed as the mean ± standard deviation (n=3). **P<0.01 vs. grade I. CREB3L1, cyclic adenosine monophosphate responsive element binding protein 3 like 1; CHOP, <t>C/EBP-homologous</t> protein; GRP78, glucose-regulated protein, 78 kDa.
Chop, 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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rabbit anti ddit3 chop  (Cell Signaling Technology Inc)
98
Cell Signaling Technology Inc rabbit anti ddit3 chop
PUS7 is a direct transcriptional target of MYC oncoproteins. A, Microarray data show mRNA expression of PUS family genes in non– MYCN -amplified neuroblastoma SK-N-AS cells with MYCN overexpression compared with vector control cells (dashed line). B and C, qRT-PCR ( B ) and immunoblotting ( C ) show MYCN-mediated upregulation of PUS7 mRNA and protein in non– MYCN -amplified neuroblastoma SK-N-AS cells with inducible MYCN overexpression in the absence of Doxy and SHEP1 cells with constitutive MYCN overexpression. PUS7 protein levels were quantified against α-tubulin (α-tub). D and E, qRT-PCR ( D ) and immunoblotting ( E ) show downregulation of PUS7 mRNA and protein expression by shRNA-mediated MYCN knockdown in MYCN -amplified neuroblastoma cell lines. PUS7 protein levels were quantified against GAPDH. F, Immunoblotting show MYC upregulation of PUS7 in P493-6 cells with inducible MYC expression in the absence of Doxy (tetoff-MYC). PUS7 protein levels were quantified against β-actin. G, ChIP-qPCR show endogenous MYCN binding to the promoter and first intron of PUS7 in MYCN -amplified neuroblastoma BE(2)-C cells. The MDM2 and <t>DDIT3</t> promoters were used as positive control for MYCN and ATF4 binding, respectively. The number associated with each primer set indicates the position of the forward primer relative to its target gene transcription start site (+1). H, Violin plot shows higher PUS7 expression in MYCN -amplified neuroblastoma tumors [the Sequencing Quality Control (SEQC) cohort, n = 498]. P values were calculated using one-way ANOVA. I, RNA-seq data show increased Pus7 mRNA expression in B cells during lymphoma development in Eµ -myc mice. PCR data ( B , D , and G ) are presented as mean ± SD ( B and D , n = 4; G , n = 3) and analyzed using two-way ANOVA. **, P < 0.01; ***, P < 0.001.
Rabbit Anti Ddit3 Chop, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 98 stars, based on 1 article reviews
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anti gadd153 chop antibody  (Novus Biologicals)
94
Novus Biologicals anti gadd153 chop antibody
PUS7 is a direct transcriptional target of MYC oncoproteins. A, Microarray data show mRNA expression of PUS family genes in non– MYCN -amplified neuroblastoma SK-N-AS cells with MYCN overexpression compared with vector control cells (dashed line). B and C, qRT-PCR ( B ) and immunoblotting ( C ) show MYCN-mediated upregulation of PUS7 mRNA and protein in non– MYCN -amplified neuroblastoma SK-N-AS cells with inducible MYCN overexpression in the absence of Doxy and SHEP1 cells with constitutive MYCN overexpression. PUS7 protein levels were quantified against α-tubulin (α-tub). D and E, qRT-PCR ( D ) and immunoblotting ( E ) show downregulation of PUS7 mRNA and protein expression by shRNA-mediated MYCN knockdown in MYCN -amplified neuroblastoma cell lines. PUS7 protein levels were quantified against GAPDH. F, Immunoblotting show MYC upregulation of PUS7 in P493-6 cells with inducible MYC expression in the absence of Doxy (tetoff-MYC). PUS7 protein levels were quantified against β-actin. G, ChIP-qPCR show endogenous MYCN binding to the promoter and first intron of PUS7 in MYCN -amplified neuroblastoma BE(2)-C cells. The MDM2 and <t>DDIT3</t> promoters were used as positive control for MYCN and ATF4 binding, respectively. The number associated with each primer set indicates the position of the forward primer relative to its target gene transcription start site (+1). H, Violin plot shows higher PUS7 expression in MYCN -amplified neuroblastoma tumors [the Sequencing Quality Control (SEQC) cohort, n = 498]. P values were calculated using one-way ANOVA. I, RNA-seq data show increased Pus7 mRNA expression in B cells during lymphoma development in Eµ -myc mice. PCR data ( B , D , and G ) are presented as mean ± SD ( B and D , n = 4; G , n = 3) and analyzed using two-way ANOVA. **, P < 0.01; ***, P < 0.001.
Anti Gadd153 Chop Antibody, supplied by Novus Biologicals, 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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anti gadd153 chop antibody - by Bioz Stars, 2026-03
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chop gadd 153 antibodies  (Santa Cruz Biotechnology)
96
Santa Cruz Biotechnology chop gadd 153 antibodies
PUS7 is a direct transcriptional target of MYC oncoproteins. A, Microarray data show mRNA expression of PUS family genes in non– MYCN -amplified neuroblastoma SK-N-AS cells with MYCN overexpression compared with vector control cells (dashed line). B and C, qRT-PCR ( B ) and immunoblotting ( C ) show MYCN-mediated upregulation of PUS7 mRNA and protein in non– MYCN -amplified neuroblastoma SK-N-AS cells with inducible MYCN overexpression in the absence of Doxy and SHEP1 cells with constitutive MYCN overexpression. PUS7 protein levels were quantified against α-tubulin (α-tub). D and E, qRT-PCR ( D ) and immunoblotting ( E ) show downregulation of PUS7 mRNA and protein expression by shRNA-mediated MYCN knockdown in MYCN -amplified neuroblastoma cell lines. PUS7 protein levels were quantified against GAPDH. F, Immunoblotting show MYC upregulation of PUS7 in P493-6 cells with inducible MYC expression in the absence of Doxy (tetoff-MYC). PUS7 protein levels were quantified against β-actin. G, ChIP-qPCR show endogenous MYCN binding to the promoter and first intron of PUS7 in MYCN -amplified neuroblastoma BE(2)-C cells. The MDM2 and <t>DDIT3</t> promoters were used as positive control for MYCN and ATF4 binding, respectively. The number associated with each primer set indicates the position of the forward primer relative to its target gene transcription start site (+1). H, Violin plot shows higher PUS7 expression in MYCN -amplified neuroblastoma tumors [the Sequencing Quality Control (SEQC) cohort, n = 498]. P values were calculated using one-way ANOVA. I, RNA-seq data show increased Pus7 mRNA expression in B cells during lymphoma development in Eµ -myc mice. PCR data ( B , D , and G ) are presented as mean ± SD ( B and D , n = 4; G , n = 3) and analyzed using two-way ANOVA. **, P < 0.01; ***, P < 0.001.
Chop Gadd 153 Antibodies, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/chop gadd 153 antibodies/product/Santa Cruz Biotechnology
Average 96 stars, based on 1 article reviews
chop gadd 153 antibodies - by Bioz Stars, 2026-03
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ab 2616025 anti chop cell signaling technology  (Cell Signaling Technology Inc)
99
Cell Signaling Technology Inc ab 2616025 anti chop cell signaling technology
PUS7 is a direct transcriptional target of MYC oncoproteins. A, Microarray data show mRNA expression of PUS family genes in non– MYCN -amplified neuroblastoma SK-N-AS cells with MYCN overexpression compared with vector control cells (dashed line). B and C, qRT-PCR ( B ) and immunoblotting ( C ) show MYCN-mediated upregulation of PUS7 mRNA and protein in non– MYCN -amplified neuroblastoma SK-N-AS cells with inducible MYCN overexpression in the absence of Doxy and SHEP1 cells with constitutive MYCN overexpression. PUS7 protein levels were quantified against α-tubulin (α-tub). D and E, qRT-PCR ( D ) and immunoblotting ( E ) show downregulation of PUS7 mRNA and protein expression by shRNA-mediated MYCN knockdown in MYCN -amplified neuroblastoma cell lines. PUS7 protein levels were quantified against GAPDH. F, Immunoblotting show MYC upregulation of PUS7 in P493-6 cells with inducible MYC expression in the absence of Doxy (tetoff-MYC). PUS7 protein levels were quantified against β-actin. G, ChIP-qPCR show endogenous MYCN binding to the promoter and first intron of PUS7 in MYCN -amplified neuroblastoma BE(2)-C cells. The MDM2 and <t>DDIT3</t> promoters were used as positive control for MYCN and ATF4 binding, respectively. The number associated with each primer set indicates the position of the forward primer relative to its target gene transcription start site (+1). H, Violin plot shows higher PUS7 expression in MYCN -amplified neuroblastoma tumors [the Sequencing Quality Control (SEQC) cohort, n = 498]. P values were calculated using one-way ANOVA. I, RNA-seq data show increased Pus7 mRNA expression in B cells during lymphoma development in Eµ -myc mice. PCR data ( B , D , and G ) are presented as mean ± SD ( B and D , n = 4; G , n = 3) and analyzed using two-way ANOVA. **, P < 0.01; ***, P < 0.001.
Ab 2616025 Anti Chop Cell Signaling Technology, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/ab 2616025 anti chop cell signaling technology/product/Cell Signaling Technology Inc
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chop d46f1 rabbit anti human monoclonal antibody  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc chop d46f1 rabbit anti human monoclonal antibody
PUS7 is a direct transcriptional target of MYC oncoproteins. A, Microarray data show mRNA expression of PUS family genes in non– MYCN -amplified neuroblastoma SK-N-AS cells with MYCN overexpression compared with vector control cells (dashed line). B and C, qRT-PCR ( B ) and immunoblotting ( C ) show MYCN-mediated upregulation of PUS7 mRNA and protein in non– MYCN -amplified neuroblastoma SK-N-AS cells with inducible MYCN overexpression in the absence of Doxy and SHEP1 cells with constitutive MYCN overexpression. PUS7 protein levels were quantified against α-tubulin (α-tub). D and E, qRT-PCR ( D ) and immunoblotting ( E ) show downregulation of PUS7 mRNA and protein expression by shRNA-mediated MYCN knockdown in MYCN -amplified neuroblastoma cell lines. PUS7 protein levels were quantified against GAPDH. F, Immunoblotting show MYC upregulation of PUS7 in P493-6 cells with inducible MYC expression in the absence of Doxy (tetoff-MYC). PUS7 protein levels were quantified against β-actin. G, ChIP-qPCR show endogenous MYCN binding to the promoter and first intron of PUS7 in MYCN -amplified neuroblastoma BE(2)-C cells. The MDM2 and <t>DDIT3</t> promoters were used as positive control for MYCN and ATF4 binding, respectively. The number associated with each primer set indicates the position of the forward primer relative to its target gene transcription start site (+1). H, Violin plot shows higher PUS7 expression in MYCN -amplified neuroblastoma tumors [the Sequencing Quality Control (SEQC) cohort, n = 498]. P values were calculated using one-way ANOVA. I, RNA-seq data show increased Pus7 mRNA expression in B cells during lymphoma development in Eµ -myc mice. PCR data ( B , D , and G ) are presented as mean ± SD ( B and D , n = 4; G , n = 3) and analyzed using two-way ANOVA. **, P < 0.01; ***, P < 0.001.
Chop D46f1 Rabbit Anti Human Monoclonal Antibody, supplied by Cell Signaling Technology Inc, 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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gene exp ddit3 rn00492098 g1  (Thermo Fisher)
92
Thermo Fisher gene exp ddit3 rn00492098 g1
PUS7 is a direct transcriptional target of MYC oncoproteins. A, Microarray data show mRNA expression of PUS family genes in non– MYCN -amplified neuroblastoma SK-N-AS cells with MYCN overexpression compared with vector control cells (dashed line). B and C, qRT-PCR ( B ) and immunoblotting ( C ) show MYCN-mediated upregulation of PUS7 mRNA and protein in non– MYCN -amplified neuroblastoma SK-N-AS cells with inducible MYCN overexpression in the absence of Doxy and SHEP1 cells with constitutive MYCN overexpression. PUS7 protein levels were quantified against α-tubulin (α-tub). D and E, qRT-PCR ( D ) and immunoblotting ( E ) show downregulation of PUS7 mRNA and protein expression by shRNA-mediated MYCN knockdown in MYCN -amplified neuroblastoma cell lines. PUS7 protein levels were quantified against GAPDH. F, Immunoblotting show MYC upregulation of PUS7 in P493-6 cells with inducible MYC expression in the absence of Doxy (tetoff-MYC). PUS7 protein levels were quantified against β-actin. G, ChIP-qPCR show endogenous MYCN binding to the promoter and first intron of PUS7 in MYCN -amplified neuroblastoma BE(2)-C cells. The MDM2 and <t>DDIT3</t> promoters were used as positive control for MYCN and ATF4 binding, respectively. The number associated with each primer set indicates the position of the forward primer relative to its target gene transcription start site (+1). H, Violin plot shows higher PUS7 expression in MYCN -amplified neuroblastoma tumors [the Sequencing Quality Control (SEQC) cohort, n = 498]. P values were calculated using one-way ANOVA. I, RNA-seq data show increased Pus7 mRNA expression in B cells during lymphoma development in Eµ -myc mice. PCR data ( B , D , and G ) are presented as mean ± SD ( B and D , n = 4; G , n = 3) and analyzed using two-way ANOVA. **, P < 0.01; ***, P < 0.001.
Gene Exp Ddit3 Rn00492098 G1, supplied by Thermo Fisher, 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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Average 92 stars, based on 1 article reviews
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ddit3 hpa068416 antibody  (Human Protein Atlas)
90
Human Protein Atlas ddit3 hpa068416 antibody
PUS7 is a direct transcriptional target of MYC oncoproteins. A, Microarray data show mRNA expression of PUS family genes in non– MYCN -amplified neuroblastoma SK-N-AS cells with MYCN overexpression compared with vector control cells (dashed line). B and C, qRT-PCR ( B ) and immunoblotting ( C ) show MYCN-mediated upregulation of PUS7 mRNA and protein in non– MYCN -amplified neuroblastoma SK-N-AS cells with inducible MYCN overexpression in the absence of Doxy and SHEP1 cells with constitutive MYCN overexpression. PUS7 protein levels were quantified against α-tubulin (α-tub). D and E, qRT-PCR ( D ) and immunoblotting ( E ) show downregulation of PUS7 mRNA and protein expression by shRNA-mediated MYCN knockdown in MYCN -amplified neuroblastoma cell lines. PUS7 protein levels were quantified against GAPDH. F, Immunoblotting show MYC upregulation of PUS7 in P493-6 cells with inducible MYC expression in the absence of Doxy (tetoff-MYC). PUS7 protein levels were quantified against β-actin. G, ChIP-qPCR show endogenous MYCN binding to the promoter and first intron of PUS7 in MYCN -amplified neuroblastoma BE(2)-C cells. The MDM2 and <t>DDIT3</t> promoters were used as positive control for MYCN and ATF4 binding, respectively. The number associated with each primer set indicates the position of the forward primer relative to its target gene transcription start site (+1). H, Violin plot shows higher PUS7 expression in MYCN -amplified neuroblastoma tumors [the Sequencing Quality Control (SEQC) cohort, n = 498]. P values were calculated using one-way ANOVA. I, RNA-seq data show increased Pus7 mRNA expression in B cells during lymphoma development in Eµ -myc mice. PCR data ( B , D , and G ) are presented as mean ± SD ( B and D , n = 4; G , n = 3) and analyzed using two-way ANOVA. **, P < 0.01; ***, P < 0.001.
Ddit3 Hpa068416 Antibody, supplied by Human Protein Atlas, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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9664 5a1 chop pmid  (Cell Signaling Technology Inc)
99
Cell Signaling Technology Inc 9664 5a1 chop pmid
PUS7 is a direct transcriptional target of MYC oncoproteins. A, Microarray data show mRNA expression of PUS family genes in non– MYCN -amplified neuroblastoma SK-N-AS cells with MYCN overexpression compared with vector control cells (dashed line). B and C, qRT-PCR ( B ) and immunoblotting ( C ) show MYCN-mediated upregulation of PUS7 mRNA and protein in non– MYCN -amplified neuroblastoma SK-N-AS cells with inducible MYCN overexpression in the absence of Doxy and SHEP1 cells with constitutive MYCN overexpression. PUS7 protein levels were quantified against α-tubulin (α-tub). D and E, qRT-PCR ( D ) and immunoblotting ( E ) show downregulation of PUS7 mRNA and protein expression by shRNA-mediated MYCN knockdown in MYCN -amplified neuroblastoma cell lines. PUS7 protein levels were quantified against GAPDH. F, Immunoblotting show MYC upregulation of PUS7 in P493-6 cells with inducible MYC expression in the absence of Doxy (tetoff-MYC). PUS7 protein levels were quantified against β-actin. G, ChIP-qPCR show endogenous MYCN binding to the promoter and first intron of PUS7 in MYCN -amplified neuroblastoma BE(2)-C cells. The MDM2 and <t>DDIT3</t> promoters were used as positive control for MYCN and ATF4 binding, respectively. The number associated with each primer set indicates the position of the forward primer relative to its target gene transcription start site (+1). H, Violin plot shows higher PUS7 expression in MYCN -amplified neuroblastoma tumors [the Sequencing Quality Control (SEQC) cohort, n = 498]. P values were calculated using one-way ANOVA. I, RNA-seq data show increased Pus7 mRNA expression in B cells during lymphoma development in Eµ -myc mice. PCR data ( B , D , and G ) are presented as mean ± SD ( B and D , n = 4; G , n = 3) and analyzed using two-way ANOVA. **, P < 0.01; ***, P < 0.001.
9664 5a1 Chop Pmid, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Expression of CREB3L1, GRP78 and CHOP in World Health Organization grade I-Ⅳ glioma tissues detected by immunohistochemistry (magnification, x400). Values are expressed as the mean ± standard deviation (n=3). **P<0.01 vs. grade I. CREB3L1, cyclic adenosine monophosphate responsive element binding protein 3 like 1; CHOP, C/EBP-homologous protein; GRP78, glucose-regulated protein, 78 kDa.

Journal: Molecular and Clinical Oncology

Article Title: Effects of endoplasmic reticulum stress-mediated CREB3L1 on apoptosis of glioma cells

doi: 10.3892/mco.2022.2516

Figure Lengend Snippet: Expression of CREB3L1, GRP78 and CHOP in World Health Organization grade I-Ⅳ glioma tissues detected by immunohistochemistry (magnification, x400). Values are expressed as the mean ± standard deviation (n=3). **P<0.01 vs. grade I. CREB3L1, cyclic adenosine monophosphate responsive element binding protein 3 like 1; CHOP, C/EBP-homologous protein; GRP78, glucose-regulated protein, 78 kDa.

Article Snippet: Sections were then blocked in 5% goat serum (cat. no. C0265; Beyotime Institute of Biotechnology) in PBS at room temperature for 30 min, followed by incubation with primary antibody against CREB3L1 (rabbit; cat. no. 11235-2-AP; 1:100 dilution; ProteinTech Group), glucose-regulated protein, 78 kDa [GRP78, also known as heat shock protein family A (Hsp70) member 5] (rabbit; Bs-1219R; 1:200 dilution; BIOSS) and C/EBP-homologous protein (CHOP) (rabbit; Bs-20669R; 1:300 dilution; BIOSS) in working solution overnight at 4˚C.

Techniques: Expressing, Immunohistochemistry, Standard Deviation, Binding Assay

Expression of CREB3L1, GRP78, CHOP and Bcl-2 following incubation with different concentrations of TG. Values are expressed as the mean ± standard deviation (n=3). *P<0.05, **P<0.01 vs. control. CREB3L1, cyclic adenosine monophosphate responsive element binding protein 3 like 1; CHOP, C/EBP-homologous protein; GRP78, glucose-regulated protein, 78 kDa; TG, thapsigargin.

Journal: Molecular and Clinical Oncology

Article Title: Effects of endoplasmic reticulum stress-mediated CREB3L1 on apoptosis of glioma cells

doi: 10.3892/mco.2022.2516

Figure Lengend Snippet: Expression of CREB3L1, GRP78, CHOP and Bcl-2 following incubation with different concentrations of TG. Values are expressed as the mean ± standard deviation (n=3). *P<0.05, **P<0.01 vs. control. CREB3L1, cyclic adenosine monophosphate responsive element binding protein 3 like 1; CHOP, C/EBP-homologous protein; GRP78, glucose-regulated protein, 78 kDa; TG, thapsigargin.

Article Snippet: Sections were then blocked in 5% goat serum (cat. no. C0265; Beyotime Institute of Biotechnology) in PBS at room temperature for 30 min, followed by incubation with primary antibody against CREB3L1 (rabbit; cat. no. 11235-2-AP; 1:100 dilution; ProteinTech Group), glucose-regulated protein, 78 kDa [GRP78, also known as heat shock protein family A (Hsp70) member 5] (rabbit; Bs-1219R; 1:200 dilution; BIOSS) and C/EBP-homologous protein (CHOP) (rabbit; Bs-20669R; 1:300 dilution; BIOSS) in working solution overnight at 4˚C.

Techniques: Expressing, Incubation, Standard Deviation, Binding Assay

PUS7 is a direct transcriptional target of MYC oncoproteins. A, Microarray data show mRNA expression of PUS family genes in non– MYCN -amplified neuroblastoma SK-N-AS cells with MYCN overexpression compared with vector control cells (dashed line). B and C, qRT-PCR ( B ) and immunoblotting ( C ) show MYCN-mediated upregulation of PUS7 mRNA and protein in non– MYCN -amplified neuroblastoma SK-N-AS cells with inducible MYCN overexpression in the absence of Doxy and SHEP1 cells with constitutive MYCN overexpression. PUS7 protein levels were quantified against α-tubulin (α-tub). D and E, qRT-PCR ( D ) and immunoblotting ( E ) show downregulation of PUS7 mRNA and protein expression by shRNA-mediated MYCN knockdown in MYCN -amplified neuroblastoma cell lines. PUS7 protein levels were quantified against GAPDH. F, Immunoblotting show MYC upregulation of PUS7 in P493-6 cells with inducible MYC expression in the absence of Doxy (tetoff-MYC). PUS7 protein levels were quantified against β-actin. G, ChIP-qPCR show endogenous MYCN binding to the promoter and first intron of PUS7 in MYCN -amplified neuroblastoma BE(2)-C cells. The MDM2 and DDIT3 promoters were used as positive control for MYCN and ATF4 binding, respectively. The number associated with each primer set indicates the position of the forward primer relative to its target gene transcription start site (+1). H, Violin plot shows higher PUS7 expression in MYCN -amplified neuroblastoma tumors [the Sequencing Quality Control (SEQC) cohort, n = 498]. P values were calculated using one-way ANOVA. I, RNA-seq data show increased Pus7 mRNA expression in B cells during lymphoma development in Eµ -myc mice. PCR data ( B , D , and G ) are presented as mean ± SD ( B and D , n = 4; G , n = 3) and analyzed using two-way ANOVA. **, P < 0.01; ***, P < 0.001.

Journal: Cancer Research

Article Title: MYC Drives mRNA Pseudouridylation to Mitigate Proliferation-Induced Cellular Stress during Cancer Development

doi: 10.1158/0008-5472.CAN-24-1102

Figure Lengend Snippet: PUS7 is a direct transcriptional target of MYC oncoproteins. A, Microarray data show mRNA expression of PUS family genes in non– MYCN -amplified neuroblastoma SK-N-AS cells with MYCN overexpression compared with vector control cells (dashed line). B and C, qRT-PCR ( B ) and immunoblotting ( C ) show MYCN-mediated upregulation of PUS7 mRNA and protein in non– MYCN -amplified neuroblastoma SK-N-AS cells with inducible MYCN overexpression in the absence of Doxy and SHEP1 cells with constitutive MYCN overexpression. PUS7 protein levels were quantified against α-tubulin (α-tub). D and E, qRT-PCR ( D ) and immunoblotting ( E ) show downregulation of PUS7 mRNA and protein expression by shRNA-mediated MYCN knockdown in MYCN -amplified neuroblastoma cell lines. PUS7 protein levels were quantified against GAPDH. F, Immunoblotting show MYC upregulation of PUS7 in P493-6 cells with inducible MYC expression in the absence of Doxy (tetoff-MYC). PUS7 protein levels were quantified against β-actin. G, ChIP-qPCR show endogenous MYCN binding to the promoter and first intron of PUS7 in MYCN -amplified neuroblastoma BE(2)-C cells. The MDM2 and DDIT3 promoters were used as positive control for MYCN and ATF4 binding, respectively. The number associated with each primer set indicates the position of the forward primer relative to its target gene transcription start site (+1). H, Violin plot shows higher PUS7 expression in MYCN -amplified neuroblastoma tumors [the Sequencing Quality Control (SEQC) cohort, n = 498]. P values were calculated using one-way ANOVA. I, RNA-seq data show increased Pus7 mRNA expression in B cells during lymphoma development in Eµ -myc mice. PCR data ( B , D , and G ) are presented as mean ± SD ( B and D , n = 4; G , n = 3) and analyzed using two-way ANOVA. **, P < 0.01; ***, P < 0.001.

Article Snippet: Proteins (20−50 μg) were separated on SDS-PAGE and transferred to nitrocellulose membranes, which were then probed with the following primary antibodies: rabbit anti-ASNS (1:1,000, Proteintech, 14681-1-AP, RRID: AB_2060119), mouse anti-ATF4 (1:400, Santa Cruz Biotechnology, sc390063, RRID: AB_2810998), rabbit anti-DENR (1:2,000, Proteintech, 10656-1-AP, RRID: AB_2092124), rabbit anti-DDIT3/CHOP (L63F6, 1:500, Cell Signaling Technology, 2895, RRID: AB_2089254), rabbit anti-GAPDH (FL335, 1:1,000, Santa Cruz Biotechnology, sc25778, RRID: AB_10167668), rabbit anti-GCN2 (1:2,000, Thermo Fisher Scientific, 720463, RRID: AB_2762413), rabbit anti-GCN2 phospho-T889 (1:1,000, Abcam, ab75836, RRID: AB_1310260), rabbit anti-MCTS1 (1:1,000, Thermo Fisher Scientific, PA5-31020, RRID: AB_2548494), rabbit anti-MYC (c-MYC, N262, 1:1,000, Santa Cruz Biotechnology, sc764, RRID: AB_631276), mouse anti-Myc tag (clone 4A6, 1:1,000, Millipore, 05724, RRID: AB_309938), mouse anti-MYCN (B8.4.B, 1:400, Santa Cruz Biotechnology, sc53993, RRID: AB_831602), rabbit anti-PHGDH (1:300, Sigma-Aldrich, HPA021241, RRID: AB_1855299), rabbit anti-PSAT1 (1:3,000, Novus Biologicals, 21020002, RRID: AB_2172599), rabbit anti-PUS7 (1:1,000, Thermo Fisher Scientific, PA5-54983, RRID: AB_2646152), rabbit anti-SLC7A5 (LAT1, 1:1,000, Cell Signaling Technology, 5347, RRID: AB_10695104), rabbit anti-SLC7A11/xCT (D2M7A, 1:1,000, Cell Signaling Technology, 12691, RRID: AB_2687474), rabbit anti-β-actin (1:2,000, Thermo Fisher Scientific, MA5-15739, RRID: AB_10979409), and mouse anti-α-tubulin (B-5-1-2, 1:5,000, Sigma-Aldrich, T5168, RRID: AB_477579).

Techniques: Microarray, Expressing, Amplification, Over Expression, Plasmid Preparation, Control, Quantitative RT-PCR, Western Blot, shRNA, Knockdown, Binding Assay, Positive Control, Sequencing, RNA Sequencing Assay