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creb3 l3  (Biorbyt)
93
Biorbyt creb3 l3
High glucose induces cAMP-responsive element-binding protein 3 Like 3 expression and the epithelial-mesenchymal transition in SV-HUC-1 cells. A: Cell Counting Kit-8 assay was employed to assess the viability of SV-HUC-1 cells following treatment with varying concentrations of glucose; B: Relative mRNA expression of cAMP-responsive element-binding protein 3 Like 3 <t>(CREB3</t> L3) in SV-HUC-1 cells after treating with different levels of glucose, as detected by quantitative PCR; C: Relative mRNA expression of CREB3 L3 in SV-HUC-1 cells with or without silencing of CREB3 L; D: SV-HUC-1 cells were treated with 5 mmol/L and 15 mmol/L glucose for 72 hours with and without CREB3 L3 knockdown (KD). Protein expression of CREB3 L3, C-reactive protein (CRP), vimentin, N-cadherin, E-cadherin, and occludin in SV-HUC-1 cells in the control (Con), high glucose (HG), HG + small interfering RNA negative control (si-NC), and HG + KD group. Each group contained three biological replicates. a P < 0.05 vs 5 mmol/L group, b P < 0.01 vs 5 mmol/L group; c P < 0.001 vs 5 mmol/L group, d P < 0.05 vs si-NC group, e P < 0.01 vs si-NC group, f P < 0.001 vs si-NC group, g P < 0.01 vs Con group, h P < 0.001 vs Con group, i P < 0.01 vs HG + si-NC group, j P < 0.001 vs HG + si-NC group.
Creb3 L3, supplied by Biorbyt, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti creb3 primary antibody  (Proteintech)
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Proteintech anti creb3 primary antibody
High glucose induces cAMP-responsive element-binding protein 3 Like 3 expression and the epithelial-mesenchymal transition in SV-HUC-1 cells. A: Cell Counting Kit-8 assay was employed to assess the viability of SV-HUC-1 cells following treatment with varying concentrations of glucose; B: Relative mRNA expression of cAMP-responsive element-binding protein 3 Like 3 <t>(CREB3</t> L3) in SV-HUC-1 cells after treating with different levels of glucose, as detected by quantitative PCR; C: Relative mRNA expression of CREB3 L3 in SV-HUC-1 cells with or without silencing of CREB3 L; D: SV-HUC-1 cells were treated with 5 mmol/L and 15 mmol/L glucose for 72 hours with and without CREB3 L3 knockdown (KD). Protein expression of CREB3 L3, C-reactive protein (CRP), vimentin, N-cadherin, E-cadherin, and occludin in SV-HUC-1 cells in the control (Con), high glucose (HG), HG + small interfering RNA negative control (si-NC), and HG + KD group. Each group contained three biological replicates. a P < 0.05 vs 5 mmol/L group, b P < 0.01 vs 5 mmol/L group; c P < 0.001 vs 5 mmol/L group, d P < 0.05 vs si-NC group, e P < 0.01 vs si-NC group, f P < 0.001 vs si-NC group, g P < 0.01 vs Con group, h P < 0.001 vs Con group, i P < 0.01 vs HG + si-NC group, j P < 0.001 vs HG + si-NC group.
Anti Creb3 Primary Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti creb3 primary antibody - by Bioz Stars, 2026-03
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rabbit polyclonal creb3 antibody  (Proteintech)
93
Proteintech rabbit polyclonal creb3 antibody
High glucose induces cAMP-responsive element-binding protein 3 Like 3 expression and the epithelial-mesenchymal transition in SV-HUC-1 cells. A: Cell Counting Kit-8 assay was employed to assess the viability of SV-HUC-1 cells following treatment with varying concentrations of glucose; B: Relative mRNA expression of cAMP-responsive element-binding protein 3 Like 3 <t>(CREB3</t> L3) in SV-HUC-1 cells after treating with different levels of glucose, as detected by quantitative PCR; C: Relative mRNA expression of CREB3 L3 in SV-HUC-1 cells with or without silencing of CREB3 L; D: SV-HUC-1 cells were treated with 5 mmol/L and 15 mmol/L glucose for 72 hours with and without CREB3 L3 knockdown (KD). Protein expression of CREB3 L3, C-reactive protein (CRP), vimentin, N-cadherin, E-cadherin, and occludin in SV-HUC-1 cells in the control (Con), high glucose (HG), HG + small interfering RNA negative control (si-NC), and HG + KD group. Each group contained three biological replicates. a P < 0.05 vs 5 mmol/L group, b P < 0.01 vs 5 mmol/L group; c P < 0.001 vs 5 mmol/L group, d P < 0.05 vs si-NC group, e P < 0.01 vs si-NC group, f P < 0.001 vs si-NC group, g P < 0.01 vs Con group, h P < 0.001 vs Con group, i P < 0.01 vs HG + si-NC group, j P < 0.001 vs HG + si-NC group.
Rabbit Polyclonal Creb3 Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit polyclonal creb3 antibody - by Bioz Stars, 2026-03
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creb3  (Proteintech)
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Proteintech creb3
The Ferroptosis-Related Differentially Expressed Genes (FRDEGs) in Degenerated Annulus Fibrosus
Creb3, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti creb3 antibody  (Danaher Inc)
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Danaher Inc anti creb3 antibody
In vitro experimental validation of <t>CREB3.</t> (A) Heatmap demonstrating top 5 TFs in different tumour cell types. (B) Western blot detects protein expression levels after CREB3 knockdown. (C, D) CCK‐8 assay showed a significant decrease in cell viability after CREB3 knockdown. (E) The colony formation experiment showed that the number of colonies of CREB3 knockdown cells was significantly lower than that of si‐NC group. (F) EdU staining assay showed that CREB3 knockdown hindered the proliferation of AGS and SGC‐7901 cells. (G) The scratch experiments showed that CREB3 knockdown significantly slowed down the migration of AGS and SGC‐7901 cells. (H) Transwell experiments showed that CREB3 knockdown significantly slowed down the invasion of AGS and SGC‐7901 cells. ** p < 0.01, *** p < 0.001.
Anti Creb3 Antibody, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit antibodies against creb3  (Proteintech)
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Proteintech rabbit antibodies against creb3
Fig. 2. Transcription factor <t>CREB3</t> binds to the ZFAS1 promoter to increase ZFAS1 expression. Notes: (A) The binding sites of CREB3 were predicted by JASPAR database. (B) GEPIA2 database predicted a prominently high expression of CREB3 gene in HCC, in which red represented HCC and grey represented the normal control, * P < 0.05 for comparison between two groups. (C) Correlation analysis between CREB3 and ZFAS1 using GEPIA2 in HCC. (D) The correlation between CREB3 and ZFAS1 in HCC was evaluated by Pearson correlation coefficient. (E) RT-qPCR and western blotting were applied to determine CREB3 expression in HCC cell lines, * P < 0.05 compared with the WT group. (F) ChIP assay was carried out to evaluate the enrichment of CREB3 protein in the ZFAS1 promoter, * P < 0.05 compared with the IgG group. (G) RT-qPCR was conducted to detect transfection efficiency, * P < 0.05 compared with the sh-NC group. (H-I) Dual-luciferase reporter gene assay was performed in Huh7 cell line after co-transfection of full-length ZFAS1 promoter or ZFAS1 promoter fragment at deletion site 1 or site 2 with sh-CREB3 vector or blank vector, * P < 0.05 compared with the sh-NC group. (J) RT-qPCR was performed to test ZFAS1 expression in Huh7 cell lines, * P < 0.05 compared with the sh-NC group. CREB3, cAMP response element binding Protein 3; ZFAS1, lncRNA Zinc Finger NFX1-Type Containing 1 antisense RNA 1; HCC, hepatocellular carcinoma.
Rabbit Antibodies Against Creb3, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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High glucose induces cAMP-responsive element-binding protein 3 Like 3 expression and the epithelial-mesenchymal transition in SV-HUC-1 cells. A: Cell Counting Kit-8 assay was employed to assess the viability of SV-HUC-1 cells following treatment with varying concentrations of glucose; B: Relative mRNA expression of cAMP-responsive element-binding protein 3 Like 3 (CREB3 L3) in SV-HUC-1 cells after treating with different levels of glucose, as detected by quantitative PCR; C: Relative mRNA expression of CREB3 L3 in SV-HUC-1 cells with or without silencing of CREB3 L; D: SV-HUC-1 cells were treated with 5 mmol/L and 15 mmol/L glucose for 72 hours with and without CREB3 L3 knockdown (KD). Protein expression of CREB3 L3, C-reactive protein (CRP), vimentin, N-cadherin, E-cadherin, and occludin in SV-HUC-1 cells in the control (Con), high glucose (HG), HG + small interfering RNA negative control (si-NC), and HG + KD group. Each group contained three biological replicates. a P < 0.05 vs 5 mmol/L group, b P < 0.01 vs 5 mmol/L group; c P < 0.001 vs 5 mmol/L group, d P < 0.05 vs si-NC group, e P < 0.01 vs si-NC group, f P < 0.001 vs si-NC group, g P < 0.01 vs Con group, h P < 0.001 vs Con group, i P < 0.01 vs HG + si-NC group, j P < 0.001 vs HG + si-NC group.

Journal: World Journal of Diabetes

Article Title: Hyperglycemia-induced overexpression of CREB3 L3 promotes the epithelial-to-mesenchymal transition in bladder urothelial cells in diabetes mellitus

doi: 10.4239/wjd.v16.i8.108101

Figure Lengend Snippet: High glucose induces cAMP-responsive element-binding protein 3 Like 3 expression and the epithelial-mesenchymal transition in SV-HUC-1 cells. A: Cell Counting Kit-8 assay was employed to assess the viability of SV-HUC-1 cells following treatment with varying concentrations of glucose; B: Relative mRNA expression of cAMP-responsive element-binding protein 3 Like 3 (CREB3 L3) in SV-HUC-1 cells after treating with different levels of glucose, as detected by quantitative PCR; C: Relative mRNA expression of CREB3 L3 in SV-HUC-1 cells with or without silencing of CREB3 L; D: SV-HUC-1 cells were treated with 5 mmol/L and 15 mmol/L glucose for 72 hours with and without CREB3 L3 knockdown (KD). Protein expression of CREB3 L3, C-reactive protein (CRP), vimentin, N-cadherin, E-cadherin, and occludin in SV-HUC-1 cells in the control (Con), high glucose (HG), HG + small interfering RNA negative control (si-NC), and HG + KD group. Each group contained three biological replicates. a P < 0.05 vs 5 mmol/L group, b P < 0.01 vs 5 mmol/L group; c P < 0.001 vs 5 mmol/L group, d P < 0.05 vs si-NC group, e P < 0.01 vs si-NC group, f P < 0.001 vs si-NC group, g P < 0.01 vs Con group, h P < 0.001 vs Con group, i P < 0.01 vs HG + si-NC group, j P < 0.001 vs HG + si-NC group.

Article Snippet: CREB3 L3 , Biorbyt , orb382910 , 49 , Rabbit , 1:500.

Techniques: Binding Assay, Expressing, Cell Counting, Real-time Polymerase Chain Reaction, Knockdown, Control, Small Interfering RNA, Negative Control

Immunofluorescence of cAMP-responsive element-binding protein 3 Like 3, C-reactive protein, E-cadherin, and occludin in urothelial cells under high glucose condition. SV-HUC-1 cells were treated with 5 mmol/L and 15 mmol/L glucose for 72 hours with and without cAMP-responsive element-binding protein 3 Like 3 (CREB3 L3) knockdown (KD). Immunofluorescence assay was conducted to detect the expression levels of CREB3 L3, C-reactive protein (CRP), E-cadherin, and occludin protein in SV-HUC-1 cells in the control (Con), high glucose (HG), HG + small interfering RNA negative control (si-NC), and HG + KD groups. Scale bar indicates 50 μm, and each group contained three replicates. a P < 0.01 vs Con group, b P < 0.001 vs Con group, c P < 0.05 vs HG + si-NC group, d P < 0.01 vs HG + si-NC group, e P < 0.001 vs HG + si-NC group.

Journal: World Journal of Diabetes

Article Title: Hyperglycemia-induced overexpression of CREB3 L3 promotes the epithelial-to-mesenchymal transition in bladder urothelial cells in diabetes mellitus

doi: 10.4239/wjd.v16.i8.108101

Figure Lengend Snippet: Immunofluorescence of cAMP-responsive element-binding protein 3 Like 3, C-reactive protein, E-cadherin, and occludin in urothelial cells under high glucose condition. SV-HUC-1 cells were treated with 5 mmol/L and 15 mmol/L glucose for 72 hours with and without cAMP-responsive element-binding protein 3 Like 3 (CREB3 L3) knockdown (KD). Immunofluorescence assay was conducted to detect the expression levels of CREB3 L3, C-reactive protein (CRP), E-cadherin, and occludin protein in SV-HUC-1 cells in the control (Con), high glucose (HG), HG + small interfering RNA negative control (si-NC), and HG + KD groups. Scale bar indicates 50 μm, and each group contained three replicates. a P < 0.01 vs Con group, b P < 0.001 vs Con group, c P < 0.05 vs HG + si-NC group, d P < 0.01 vs HG + si-NC group, e P < 0.001 vs HG + si-NC group.

Article Snippet: CREB3 L3 , Biorbyt , orb382910 , 49 , Rabbit , 1:500.

Techniques: Immunofluorescence, Binding Assay, Knockdown, Expressing, Control, Small Interfering RNA, Negative Control

cAMP-responsive element-binding protein 3 Like 3 upregulates C-reactive protein expression, induces the epithelial-mesenchymal transition, and impairs cellular tight junctions in the diabetic cystopathy rat model. A: Western blot analysis of cAMP-responsive element-binding protein 3 Like 3 (CREB3 L3), C-reactive protein (CRP), vimentin, N-cadherin, E-cadherin, claudin 1, and occludin in the bladder urothelial tissues from each group; B: Immunofluorescence analysis of the expression of CREB3 L3, N-cadherin, E-cadherin, claudin 1, and occludin proteins in the bladder urothelial tissues from each group. Each group contained three biological replicates. Scale bar indicates 50 μm. a P < 0.05 vs negative control (NC) group, b P < 0.01 vs NC group, c P < 0.001 vs NC group; d P < 0.05 vs diabetes mellitus (DM) group, e P < 0.01 vs DM group, f P < 0.001 vs DM group.

Journal: World Journal of Diabetes

Article Title: Hyperglycemia-induced overexpression of CREB3 L3 promotes the epithelial-to-mesenchymal transition in bladder urothelial cells in diabetes mellitus

doi: 10.4239/wjd.v16.i8.108101

Figure Lengend Snippet: cAMP-responsive element-binding protein 3 Like 3 upregulates C-reactive protein expression, induces the epithelial-mesenchymal transition, and impairs cellular tight junctions in the diabetic cystopathy rat model. A: Western blot analysis of cAMP-responsive element-binding protein 3 Like 3 (CREB3 L3), C-reactive protein (CRP), vimentin, N-cadherin, E-cadherin, claudin 1, and occludin in the bladder urothelial tissues from each group; B: Immunofluorescence analysis of the expression of CREB3 L3, N-cadherin, E-cadherin, claudin 1, and occludin proteins in the bladder urothelial tissues from each group. Each group contained three biological replicates. Scale bar indicates 50 μm. a P < 0.05 vs negative control (NC) group, b P < 0.01 vs NC group, c P < 0.001 vs NC group; d P < 0.05 vs diabetes mellitus (DM) group, e P < 0.01 vs DM group, f P < 0.001 vs DM group.

Article Snippet: CREB3 L3 , Biorbyt , orb382910 , 49 , Rabbit , 1:500.

Techniques: Binding Assay, Expressing, Western Blot, Immunofluorescence, Negative Control

Establishment of diabetic cystopathy rat models for 8 weeks. A: At the end of the 8 weeks, the body weight, blood glucose and bladder weight from rats were measured; B: Hematoxylin and eosin staining of bladder tissues of the rats at the end of the 8 weeks, and each group contained three replicates; C: Western blotting detected the expression of cAMP-responsive element-binding protein 3 Like 3 (CREB3 L3), C-reactive protein (CRP), vimentin, N-cadherin, E-cadherin, claudin 1, and occludin in the bladder urothelial tissues of the rats at the end of 8 weeks. Each group contained three replicates. a P < 0.05 vs negative control (NC) group, b P < 0.01 vs NC group, c P < 0.001 vs NC group; d P < 0.05 vs diabetes mellitus (DM) group, e P < 0.01 vs DM group, f P < 0.001 vs DM group.

Journal: World Journal of Diabetes

Article Title: Hyperglycemia-induced overexpression of CREB3 L3 promotes the epithelial-to-mesenchymal transition in bladder urothelial cells in diabetes mellitus

doi: 10.4239/wjd.v16.i8.108101

Figure Lengend Snippet: Establishment of diabetic cystopathy rat models for 8 weeks. A: At the end of the 8 weeks, the body weight, blood glucose and bladder weight from rats were measured; B: Hematoxylin and eosin staining of bladder tissues of the rats at the end of the 8 weeks, and each group contained three replicates; C: Western blotting detected the expression of cAMP-responsive element-binding protein 3 Like 3 (CREB3 L3), C-reactive protein (CRP), vimentin, N-cadherin, E-cadherin, claudin 1, and occludin in the bladder urothelial tissues of the rats at the end of 8 weeks. Each group contained three replicates. a P < 0.05 vs negative control (NC) group, b P < 0.01 vs NC group, c P < 0.001 vs NC group; d P < 0.05 vs diabetes mellitus (DM) group, e P < 0.01 vs DM group, f P < 0.001 vs DM group.

Article Snippet: CREB3 L3 , Biorbyt , orb382910 , 49 , Rabbit , 1:500.

Techniques: Staining, Western Blot, Expressing, Binding Assay, Negative Control

cAMP-responsive element-binding protein 3 Like 3 continuously induces the epithelial-mesenchymal transition and impairs cellular tight junctions with the progression of diabetic cystopathy. At the end of 8 weeks, immunofluorescence staining of cAMP-responsive element-binding protein 3 Like 3 (CREB3 L3), N-cadherin, E-cadherin, claudin 1, and occludin proteins in the bladder urothelial tissues from each group. Each group contained three replicates. Scale bar indicates 50 μm. a P < 0.05 vs negative control (NC) group, b P < 0.01 vs NC group, c P < 0.001 vs NC group, d P < 0.0001 vs NC group; e P < 0.05 vs diabetes mellitus (DM) group, f P < 0.01 vs DM group, g P < 0.001 vs DM group.

Journal: World Journal of Diabetes

Article Title: Hyperglycemia-induced overexpression of CREB3 L3 promotes the epithelial-to-mesenchymal transition in bladder urothelial cells in diabetes mellitus

doi: 10.4239/wjd.v16.i8.108101

Figure Lengend Snippet: cAMP-responsive element-binding protein 3 Like 3 continuously induces the epithelial-mesenchymal transition and impairs cellular tight junctions with the progression of diabetic cystopathy. At the end of 8 weeks, immunofluorescence staining of cAMP-responsive element-binding protein 3 Like 3 (CREB3 L3), N-cadherin, E-cadherin, claudin 1, and occludin proteins in the bladder urothelial tissues from each group. Each group contained three replicates. Scale bar indicates 50 μm. a P < 0.05 vs negative control (NC) group, b P < 0.01 vs NC group, c P < 0.001 vs NC group, d P < 0.0001 vs NC group; e P < 0.05 vs diabetes mellitus (DM) group, f P < 0.01 vs DM group, g P < 0.001 vs DM group.

Article Snippet: CREB3 L3 , Biorbyt , orb382910 , 49 , Rabbit , 1:500.

Techniques: Binding Assay, Immunofluorescence, Staining, Negative Control

Establishment of diabetic cystopathy models of rats for 12 weeks. A: At the end of the 12 weeks, the body weight, blood glucose and bladder weight from rats were measured; B: Hematoxylin and eosin staining of bladder tissues from rats at the end of the 8 and 12 weeks; C: Western blotting detected the expression of cAMP-responsive element-binding protein 3 Like 3 (CREB3 L3), C-reactive protein (CRP), vimentin, N-cadherin, E-cadherin, claudin 1, and occludin proteins in the bladder urothelial tissues from each group at the end of 12 weeks. Each group contained three replicates. a P < 0.01 vs negative control (NC) group, b P < 0.001 vs NC group; c P < 0.01 vs diabetes mellitus (DM) group, d P < 0.001 vs DM group.

Journal: World Journal of Diabetes

Article Title: Hyperglycemia-induced overexpression of CREB3 L3 promotes the epithelial-to-mesenchymal transition in bladder urothelial cells in diabetes mellitus

doi: 10.4239/wjd.v16.i8.108101

Figure Lengend Snippet: Establishment of diabetic cystopathy models of rats for 12 weeks. A: At the end of the 12 weeks, the body weight, blood glucose and bladder weight from rats were measured; B: Hematoxylin and eosin staining of bladder tissues from rats at the end of the 8 and 12 weeks; C: Western blotting detected the expression of cAMP-responsive element-binding protein 3 Like 3 (CREB3 L3), C-reactive protein (CRP), vimentin, N-cadherin, E-cadherin, claudin 1, and occludin proteins in the bladder urothelial tissues from each group at the end of 12 weeks. Each group contained three replicates. a P < 0.01 vs negative control (NC) group, b P < 0.001 vs NC group; c P < 0.01 vs diabetes mellitus (DM) group, d P < 0.001 vs DM group.

Article Snippet: CREB3 L3 , Biorbyt , orb382910 , 49 , Rabbit , 1:500.

Techniques: Staining, Western Blot, Expressing, Binding Assay, Negative Control

cAMP-responsive element-binding protein 3 Like 3 results in the epithelial-mesenchymal transition and impairs cellular tight junctions in the late stage of diabetic cystopathy. At the end of 12 weeks, immunofluorescence staining of cAMP-responsive element-binding protein 3 Like 3 (CREB3 L3), N-cadherin, E-cadherin, claudin 1, and occludin proteins in the bladder urothelial tissues from each group. Each group contained three biological replicates. Scale bar indicates 50 μm. a P < 0.01 vs negative control (NC) group, b P < 0.001 vs NC group, c P < 0.0001 vs NC group; d P < 0.05 vs diabetes mellitus (DM) group, e P < 0.01 vs DM group.

Journal: World Journal of Diabetes

Article Title: Hyperglycemia-induced overexpression of CREB3 L3 promotes the epithelial-to-mesenchymal transition in bladder urothelial cells in diabetes mellitus

doi: 10.4239/wjd.v16.i8.108101

Figure Lengend Snippet: cAMP-responsive element-binding protein 3 Like 3 results in the epithelial-mesenchymal transition and impairs cellular tight junctions in the late stage of diabetic cystopathy. At the end of 12 weeks, immunofluorescence staining of cAMP-responsive element-binding protein 3 Like 3 (CREB3 L3), N-cadherin, E-cadherin, claudin 1, and occludin proteins in the bladder urothelial tissues from each group. Each group contained three biological replicates. Scale bar indicates 50 μm. a P < 0.01 vs negative control (NC) group, b P < 0.001 vs NC group, c P < 0.0001 vs NC group; d P < 0.05 vs diabetes mellitus (DM) group, e P < 0.01 vs DM group.

Article Snippet: CREB3 L3 , Biorbyt , orb382910 , 49 , Rabbit , 1:500.

Techniques: Binding Assay, Immunofluorescence, Staining, Negative Control

The Ferroptosis-Related Differentially Expressed Genes (FRDEGs) in Degenerated Annulus Fibrosus

Journal: Journal of Inflammation Research

Article Title: MUC1 and CREB3 are Hub Ferroptosis Suppressors for Nucleus Pulposus and Annulus Fibrosus Degeneration by Integrated Bioinformatics and Experimental Verification

doi: 10.2147/JIR.S489052

Figure Lengend Snippet: The Ferroptosis-Related Differentially Expressed Genes (FRDEGs) in Degenerated Annulus Fibrosus

Article Snippet: Cells were fixed with 4% paraformaldehyde solution, then permeabilized with 0.1% Triton-X100 solution for 30 min. After blocking with 1% goat serum, the cells were incubated overnight at 4°C with rabbit-derived MUC1 (Abcam, ab109185) and CREB3 (Proteintech, 11275-1-AP) antibody (diluted in 1% BSA-PBS at a dilution ratio of 1:200).

Techniques: Expressing, Marker

The expression level of each hub gene in the validation set. ( A ) Principal component analysis (PCA) of all the nucleus pulposus (NP) samples in GSE167199, GSE186542, and GSE244889. The expression of AKR1C1 ( B ), AKR1C3 ( C ), MUC1 ( D ), and ENPP2 ( E ) in NP samples. The expression of SCP2 ( F ), ABCC1 ( G ), KLF2 ( H ), IDO1 ( I ), and CREB3 ( J ) in annulus fibrosus samples. *p < 0.05; ns, nonstatistical significance.

Journal: Journal of Inflammation Research

Article Title: MUC1 and CREB3 are Hub Ferroptosis Suppressors for Nucleus Pulposus and Annulus Fibrosus Degeneration by Integrated Bioinformatics and Experimental Verification

doi: 10.2147/JIR.S489052

Figure Lengend Snippet: The expression level of each hub gene in the validation set. ( A ) Principal component analysis (PCA) of all the nucleus pulposus (NP) samples in GSE167199, GSE186542, and GSE244889. The expression of AKR1C1 ( B ), AKR1C3 ( C ), MUC1 ( D ), and ENPP2 ( E ) in NP samples. The expression of SCP2 ( F ), ABCC1 ( G ), KLF2 ( H ), IDO1 ( I ), and CREB3 ( J ) in annulus fibrosus samples. *p < 0.05; ns, nonstatistical significance.

Article Snippet: Cells were fixed with 4% paraformaldehyde solution, then permeabilized with 0.1% Triton-X100 solution for 30 min. After blocking with 1% goat serum, the cells were incubated overnight at 4°C with rabbit-derived MUC1 (Abcam, ab109185) and CREB3 (Proteintech, 11275-1-AP) antibody (diluted in 1% BSA-PBS at a dilution ratio of 1:200).

Techniques: Expressing, Biomarker Discovery

Single-cell RNC-seq analysis of the annulus fibrosus (AF) samples. ( A ) Visualization of clustering by TSNE plot using 0.5 resolution. ( B ) Bubble plot showing the expression of the ACAN, SOX9, and COL1A1 related different clusters. ( C ) Six cell types were identified, including AF cell (AFC), fibroblast, macrophage, muscle stem cell (MSC), nucleus pulposus cell (NPC), and T cell. ( D ) Seven subclusters for the AF cells were identified using 0.4 resolution. ( E-G ) Monocle pseudotime trajectory showing the progression of seven subclusters of AF cells. ( H ) TSNE image of CREB3 in subcluster 3 and 5, showing the high expression in cluster 3.

Journal: Journal of Inflammation Research

Article Title: MUC1 and CREB3 are Hub Ferroptosis Suppressors for Nucleus Pulposus and Annulus Fibrosus Degeneration by Integrated Bioinformatics and Experimental Verification

doi: 10.2147/JIR.S489052

Figure Lengend Snippet: Single-cell RNC-seq analysis of the annulus fibrosus (AF) samples. ( A ) Visualization of clustering by TSNE plot using 0.5 resolution. ( B ) Bubble plot showing the expression of the ACAN, SOX9, and COL1A1 related different clusters. ( C ) Six cell types were identified, including AF cell (AFC), fibroblast, macrophage, muscle stem cell (MSC), nucleus pulposus cell (NPC), and T cell. ( D ) Seven subclusters for the AF cells were identified using 0.4 resolution. ( E-G ) Monocle pseudotime trajectory showing the progression of seven subclusters of AF cells. ( H ) TSNE image of CREB3 in subcluster 3 and 5, showing the high expression in cluster 3.

Article Snippet: Cells were fixed with 4% paraformaldehyde solution, then permeabilized with 0.1% Triton-X100 solution for 30 min. After blocking with 1% goat serum, the cells were incubated overnight at 4°C with rabbit-derived MUC1 (Abcam, ab109185) and CREB3 (Proteintech, 11275-1-AP) antibody (diluted in 1% BSA-PBS at a dilution ratio of 1:200).

Techniques: Expressing

Experimental validations of MUC1 and CREB3 in nucleus pulposus (NP) and annulus fibrosus (AF). ( A and B ) Immunohistochemical results demonstrated that the expression of MUC1 was decreased in degenerated NP tissues. ( C and D ) Immunohistochemical results showed that the expression of MUC1 decreased in degenerated rat NP tissues. ( E ) Immunofluorescence detection of protein expression of MUC1 in NP cells, scale bar=50 μm. ( F and G ) Immunohistochemical results of the expression of CREB3 in degenerated rat AF tissues. The Fe 2+ ( H and J , scale bar = 20μm) and ROS ( H and K , scale bar = 100μm) intensity was increased in TBHP-treated AF cells, and mitochondrial morphology detection by transmission electron microscopy (TEM) ( I , scale bar = 1000 nm, the red arrows indicate mitochondria). ( L ) Immunofluorescence detection of protein expression of CREB3 in AF cells, scale bar=50 μm. ***p < 0.001.

Journal: Journal of Inflammation Research

Article Title: MUC1 and CREB3 are Hub Ferroptosis Suppressors for Nucleus Pulposus and Annulus Fibrosus Degeneration by Integrated Bioinformatics and Experimental Verification

doi: 10.2147/JIR.S489052

Figure Lengend Snippet: Experimental validations of MUC1 and CREB3 in nucleus pulposus (NP) and annulus fibrosus (AF). ( A and B ) Immunohistochemical results demonstrated that the expression of MUC1 was decreased in degenerated NP tissues. ( C and D ) Immunohistochemical results showed that the expression of MUC1 decreased in degenerated rat NP tissues. ( E ) Immunofluorescence detection of protein expression of MUC1 in NP cells, scale bar=50 μm. ( F and G ) Immunohistochemical results of the expression of CREB3 in degenerated rat AF tissues. The Fe 2+ ( H and J , scale bar = 20μm) and ROS ( H and K , scale bar = 100μm) intensity was increased in TBHP-treated AF cells, and mitochondrial morphology detection by transmission electron microscopy (TEM) ( I , scale bar = 1000 nm, the red arrows indicate mitochondria). ( L ) Immunofluorescence detection of protein expression of CREB3 in AF cells, scale bar=50 μm. ***p < 0.001.

Article Snippet: Cells were fixed with 4% paraformaldehyde solution, then permeabilized with 0.1% Triton-X100 solution for 30 min. After blocking with 1% goat serum, the cells were incubated overnight at 4°C with rabbit-derived MUC1 (Abcam, ab109185) and CREB3 (Proteintech, 11275-1-AP) antibody (diluted in 1% BSA-PBS at a dilution ratio of 1:200).

Techniques: Immunohistochemical staining, Expressing, Immunofluorescence, Transmission Assay, Electron Microscopy

In vitro experimental validation of CREB3. (A) Heatmap demonstrating top 5 TFs in different tumour cell types. (B) Western blot detects protein expression levels after CREB3 knockdown. (C, D) CCK‐8 assay showed a significant decrease in cell viability after CREB3 knockdown. (E) The colony formation experiment showed that the number of colonies of CREB3 knockdown cells was significantly lower than that of si‐NC group. (F) EdU staining assay showed that CREB3 knockdown hindered the proliferation of AGS and SGC‐7901 cells. (G) The scratch experiments showed that CREB3 knockdown significantly slowed down the migration of AGS and SGC‐7901 cells. (H) Transwell experiments showed that CREB3 knockdown significantly slowed down the invasion of AGS and SGC‐7901 cells. ** p < 0.01, *** p < 0.001.

Journal: Journal of Cellular and Molecular Medicine

Article Title: Single‐cell analysis uncovers high‐proliferative tumour cell subtypes and their interactions in the microenvironment of gastric cancer

doi: 10.1111/jcmm.18373

Figure Lengend Snippet: In vitro experimental validation of CREB3. (A) Heatmap demonstrating top 5 TFs in different tumour cell types. (B) Western blot detects protein expression levels after CREB3 knockdown. (C, D) CCK‐8 assay showed a significant decrease in cell viability after CREB3 knockdown. (E) The colony formation experiment showed that the number of colonies of CREB3 knockdown cells was significantly lower than that of si‐NC group. (F) EdU staining assay showed that CREB3 knockdown hindered the proliferation of AGS and SGC‐7901 cells. (G) The scratch experiments showed that CREB3 knockdown significantly slowed down the migration of AGS and SGC‐7901 cells. (H) Transwell experiments showed that CREB3 knockdown significantly slowed down the invasion of AGS and SGC‐7901 cells. ** p < 0.01, *** p < 0.001.

Article Snippet: After that, proteins were transferred to PVDF membranes, blotted with 5% BSA for 1.5 h at room temperature, incubated with Anti‐CREB3 antibody (ab180119, abcam) at 4°C overnight and then incubated with the corresponding secondary antibody for 1 h. The membranes were visualized by ECL Western Blot substrate.

Techniques: In Vitro, Western Blot, Expressing, CCK-8 Assay, Staining, Migration

Fig. 2. Transcription factor CREB3 binds to the ZFAS1 promoter to increase ZFAS1 expression. Notes: (A) The binding sites of CREB3 were predicted by JASPAR database. (B) GEPIA2 database predicted a prominently high expression of CREB3 gene in HCC, in which red represented HCC and grey represented the normal control, * P < 0.05 for comparison between two groups. (C) Correlation analysis between CREB3 and ZFAS1 using GEPIA2 in HCC. (D) The correlation between CREB3 and ZFAS1 in HCC was evaluated by Pearson correlation coefficient. (E) RT-qPCR and western blotting were applied to determine CREB3 expression in HCC cell lines, * P < 0.05 compared with the WT group. (F) ChIP assay was carried out to evaluate the enrichment of CREB3 protein in the ZFAS1 promoter, * P < 0.05 compared with the IgG group. (G) RT-qPCR was conducted to detect transfection efficiency, * P < 0.05 compared with the sh-NC group. (H-I) Dual-luciferase reporter gene assay was performed in Huh7 cell line after co-transfection of full-length ZFAS1 promoter or ZFAS1 promoter fragment at deletion site 1 or site 2 with sh-CREB3 vector or blank vector, * P < 0.05 compared with the sh-NC group. (J) RT-qPCR was performed to test ZFAS1 expression in Huh7 cell lines, * P < 0.05 compared with the sh-NC group. CREB3, cAMP response element binding Protein 3; ZFAS1, lncRNA Zinc Finger NFX1-Type Containing 1 antisense RNA 1; HCC, hepatocellular carcinoma.

Journal: Translational oncology

Article Title: CREB3 facilitates Donafenib resistance in hepatocellular carcinoma cells via the LSD1/CoREST/p65 axis by transcriptionally activating long noncoding RNA ZFAS1.

doi: 10.1016/j.tranon.2023.101684

Figure Lengend Snippet: Fig. 2. Transcription factor CREB3 binds to the ZFAS1 promoter to increase ZFAS1 expression. Notes: (A) The binding sites of CREB3 were predicted by JASPAR database. (B) GEPIA2 database predicted a prominently high expression of CREB3 gene in HCC, in which red represented HCC and grey represented the normal control, * P < 0.05 for comparison between two groups. (C) Correlation analysis between CREB3 and ZFAS1 using GEPIA2 in HCC. (D) The correlation between CREB3 and ZFAS1 in HCC was evaluated by Pearson correlation coefficient. (E) RT-qPCR and western blotting were applied to determine CREB3 expression in HCC cell lines, * P < 0.05 compared with the WT group. (F) ChIP assay was carried out to evaluate the enrichment of CREB3 protein in the ZFAS1 promoter, * P < 0.05 compared with the IgG group. (G) RT-qPCR was conducted to detect transfection efficiency, * P < 0.05 compared with the sh-NC group. (H-I) Dual-luciferase reporter gene assay was performed in Huh7 cell line after co-transfection of full-length ZFAS1 promoter or ZFAS1 promoter fragment at deletion site 1 or site 2 with sh-CREB3 vector or blank vector, * P < 0.05 compared with the sh-NC group. (J) RT-qPCR was performed to test ZFAS1 expression in Huh7 cell lines, * P < 0.05 compared with the sh-NC group. CREB3, cAMP response element binding Protein 3; ZFAS1, lncRNA Zinc Finger NFX1-Type Containing 1 antisense RNA 1; HCC, hepatocellular carcinoma.

Article Snippet: For the experimental groups, the supernatants were mixed upside down with 1 μL rabbit antibodies against CREB3 (11,275–1-AP, Proteintech, Rosemont, IL, USA), LSD1 (ab129195, Abcam), histone 3 lysine 4 monomethylation (H3K4me1, ab176877, Abcam), H3K4 dimethylation (H3K4me2, ab32356, Abcam), or IgG (ab172730, Abcam, as the NC) and 60 μL Protein A Agarose/Salmon Sperm DNA at 4 ◦C for 2 h, allowed to stand for 10 min, and centrifuged at 700 g for 1 min.

Techniques: Expressing, Binding Assay, Control, Comparison, Quantitative RT-PCR, Western Blot, Transfection, Luciferase, Reporter Gene Assay, Cotransfection, Plasmid Preparation

Fig. 3. ZFAS1 enhances p65 expression by interacting with LSD1/CoREST. Notes: (A) The correlation between ZFAS1 and p65 in HCC was analysed via Pearson correlation coefficient. (B) RT-qPCR was implemented to assess CREB3 expression in HCC cell lines, * P < 0.05 compared with the WT group. (C) Western blotting was employed to test CREB3 expression in HCC cell lines, * P < 0.05 compared with the WT group. (D) The interaction between ZFAS1 and LSD1/CoREST was assessed by RIP assay. (E) After knockdown of ZFAS1 or CREB3, ChIP assay was adopted to measure the enrichment of LSD1, H3K4me1, and H3K4me2 in p65 promoter, * P < 0.05 compared with the anti-IgG group. (F) Dual-luciferase reporter gene assay was employed to validate the binding relationship between CREB3/ZFAS1 and p65, * P < 0.05 compared with the sh-NC group. (G) RT- qPCR was carried out to assess p65 expression in Huh7 cells, * P < 0.05 compared with the sh-NC group. (H) Western blotting was utilized to determine p65 expression in Huh7 cells, * P < 0.05 compared with the sh-NC group. CREB3, cAMP response element binding Protein 3; ZFAS1, lncRNA Zinc Finger NFX1-Type Containing 1 antisense RNA 1; HCC, hepatocellular carcinoma; LSD1, lysine-specific demethylase 1; CoREST, corepressor of repressor element-1 silencing tran scription factor; H3K4me1, histone 3 lysine 4 monomethylation.

Journal: Translational oncology

Article Title: CREB3 facilitates Donafenib resistance in hepatocellular carcinoma cells via the LSD1/CoREST/p65 axis by transcriptionally activating long noncoding RNA ZFAS1.

doi: 10.1016/j.tranon.2023.101684

Figure Lengend Snippet: Fig. 3. ZFAS1 enhances p65 expression by interacting with LSD1/CoREST. Notes: (A) The correlation between ZFAS1 and p65 in HCC was analysed via Pearson correlation coefficient. (B) RT-qPCR was implemented to assess CREB3 expression in HCC cell lines, * P < 0.05 compared with the WT group. (C) Western blotting was employed to test CREB3 expression in HCC cell lines, * P < 0.05 compared with the WT group. (D) The interaction between ZFAS1 and LSD1/CoREST was assessed by RIP assay. (E) After knockdown of ZFAS1 or CREB3, ChIP assay was adopted to measure the enrichment of LSD1, H3K4me1, and H3K4me2 in p65 promoter, * P < 0.05 compared with the anti-IgG group. (F) Dual-luciferase reporter gene assay was employed to validate the binding relationship between CREB3/ZFAS1 and p65, * P < 0.05 compared with the sh-NC group. (G) RT- qPCR was carried out to assess p65 expression in Huh7 cells, * P < 0.05 compared with the sh-NC group. (H) Western blotting was utilized to determine p65 expression in Huh7 cells, * P < 0.05 compared with the sh-NC group. CREB3, cAMP response element binding Protein 3; ZFAS1, lncRNA Zinc Finger NFX1-Type Containing 1 antisense RNA 1; HCC, hepatocellular carcinoma; LSD1, lysine-specific demethylase 1; CoREST, corepressor of repressor element-1 silencing tran scription factor; H3K4me1, histone 3 lysine 4 monomethylation.

Article Snippet: For the experimental groups, the supernatants were mixed upside down with 1 μL rabbit antibodies against CREB3 (11,275–1-AP, Proteintech, Rosemont, IL, USA), LSD1 (ab129195, Abcam), histone 3 lysine 4 monomethylation (H3K4me1, ab176877, Abcam), H3K4 dimethylation (H3K4me2, ab32356, Abcam), or IgG (ab172730, Abcam, as the NC) and 60 μL Protein A Agarose/Salmon Sperm DNA at 4 ◦C for 2 h, allowed to stand for 10 min, and centrifuged at 700 g for 1 min.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Knockdown, Luciferase, Reporter Gene Assay, Binding Assay

Fig. 4. CREB3 upregulates p65 expression to enhance Donafenib resistance in HCC cells. Notes: HepG2-DR and Huh7-DR cells were transfected with sh-CREB3 and/or oe-p65. (A) CREB3, ZFAS1, and p65 expression was measured using RT-qPCR. (B) CREB3 and p65 expression was detected using western blotting. Stably transfected HepG2-DR and Huh7-DR cells were treated with Donafenib. (C) The colony formation rate of cells was assessed using colony formation assay. (D) Flow cytometry was applied to examine the apoptosis of cells. (E) MTT was adopted to determine the proliferation and IC50 of cells. * P < 0.05 compared with the sh-NC + oe-NC group; # P < 0.05 compared with the sh-CREB3 + oe-NC group. CREB3, cAMP response element binding Protein 3; ZFAS1, lncRNA Zinc Finger NFX1-Type Containing 1 antisense RNA 1; HCC, hepatocellular carcinoma; DR, Donafe nib-resistant.

Journal: Translational oncology

Article Title: CREB3 facilitates Donafenib resistance in hepatocellular carcinoma cells via the LSD1/CoREST/p65 axis by transcriptionally activating long noncoding RNA ZFAS1.

doi: 10.1016/j.tranon.2023.101684

Figure Lengend Snippet: Fig. 4. CREB3 upregulates p65 expression to enhance Donafenib resistance in HCC cells. Notes: HepG2-DR and Huh7-DR cells were transfected with sh-CREB3 and/or oe-p65. (A) CREB3, ZFAS1, and p65 expression was measured using RT-qPCR. (B) CREB3 and p65 expression was detected using western blotting. Stably transfected HepG2-DR and Huh7-DR cells were treated with Donafenib. (C) The colony formation rate of cells was assessed using colony formation assay. (D) Flow cytometry was applied to examine the apoptosis of cells. (E) MTT was adopted to determine the proliferation and IC50 of cells. * P < 0.05 compared with the sh-NC + oe-NC group; # P < 0.05 compared with the sh-CREB3 + oe-NC group. CREB3, cAMP response element binding Protein 3; ZFAS1, lncRNA Zinc Finger NFX1-Type Containing 1 antisense RNA 1; HCC, hepatocellular carcinoma; DR, Donafe nib-resistant.

Article Snippet: For the experimental groups, the supernatants were mixed upside down with 1 μL rabbit antibodies against CREB3 (11,275–1-AP, Proteintech, Rosemont, IL, USA), LSD1 (ab129195, Abcam), histone 3 lysine 4 monomethylation (H3K4me1, ab176877, Abcam), H3K4 dimethylation (H3K4me2, ab32356, Abcam), or IgG (ab172730, Abcam, as the NC) and 60 μL Protein A Agarose/Salmon Sperm DNA at 4 ◦C for 2 h, allowed to stand for 10 min, and centrifuged at 700 g for 1 min.

Techniques: Expressing, Transfection, Quantitative RT-PCR, Western Blot, Stable Transfection, Colony Assay, Flow Cytometry, Binding Assay