Journal: Biochemical Journal

Article Title: Induced JunD in intestinal epithelial cells represses CDK4 transcription through its proximal promoter region following polyamine depletion

doi: 10.1042/bj20061436

Figure Lengend Snippet: Figure 4 Effect of ectopic expression of the junD gene on CDK4 transcription

Article Snippet: The plasmid clone (pRSV-hjD) containing the human junD gene was obtained from ATCC.

Techniques: Expressing

Journal: Biochemical Journal

Article Title: Induced JunD in intestinal epithelial cells represses CDK4 transcription through its proximal promoter region following polyamine depletion

doi: 10.1042/bj20061436

Figure Lengend Snippet: Figure 5 Effect of ectopic expression of the junD gene on CDK4-promoter activity after deletion mutation of AP-1 binding site

Article Snippet: The plasmid clone (pRSV-hjD) containing the human junD gene was obtained from ATCC.

Techniques: Expressing, Activity Assay, Mutagenesis, Binding Assay

Journal: Biochemical Journal

Article Title: Induced JunD in intestinal epithelial cells represses CDK4 transcription through its proximal promoter region following polyamine depletion

doi: 10.1042/bj20061436

Figure Lengend Snippet: Figure 7 Effect of ectopic expression of the junD gene on CDK4-promoter luciferase reporter activity: AP-1 point mutation within CDK4-promoter

Article Snippet: The plasmid clone (pRSV-hjD) containing the human junD gene was obtained from ATCC.

Techniques: Expressing, Luciferase, Activity Assay, Mutagenesis

Journal: Journal of cell science

Article Title: CREB regulates the expression of type 1 inositol 1,4,5-trisphosphate receptors.

doi: 10.1242/jcs.258875

Figure Lengend Snippet: Fig. 3. Evidence for involvement of PKA–CREB axis in governing the expression of IP3R1. (A,C,E) HEK-293 cells were not transfected (0 μg, black), or transiently transfected with 0.5, 1.0 or 2.0 μg (blue, green or red, respectively) amounts of VP16-CREB (A), KCREB (C) or PKI (E) expression plasmid; total protein amounts were isolated 36 h following transfection and analyzed by western blotting. Representative western blots are shown. Overexpression of VP16- CREB resulted in a dose-dependent increase in levels of endogenous IP3R1 (A). Blocking endogenous CREB due to overexpression of KCREB resulted in a dose-dependent decrease of endogenous IP3R1 (C). Inhibition of endogenous PKA due to overexpression of PKI, resulted in a dose-dependent decrease of endogenous IP3R1 (E). (B,D,F) Quantitative analysis of three independent western blots for experimental scenarios described under A, C and E. Statistical significance was determined by one-way ANOVA with Tukey’s multiple comparisons post-test. *P<0.05, **P<0.01. (G,H) HEK-293 cells were transiently transfected (+) with either pcDNA control or KCREB plasmid. After 24 h, cells were further treated with 10 μM forskolin (+) or left untreated (−) for an additional 12 h. At 36 h after transfection, protein levels were analyzed by western blotting, showing increased levels of IP3R1 in cells treated with forskolin under basal conditions or when CREB was inhibited using KCREB, suggesting involvement of CREB in forskolin-induced expression of IP3R1. (H) Quantitative analysis of three independent western blots for the experimental scenario described under G. Statistical significance was determined by Student’s t-test (unpaired, two- tailed). *P<0.05 with respect to control condition. ns, not significant.

Article Snippet: ITPR2 promoterreporter construct was kindly provided by Michael Nathanson (Yale University, New Haven, CT). pRL-TK plasmid was kindly provided by Cesare Orlandi (University of Rochester, Rochester, NY). pRSV-PKI-v2, expressing the PKA inhibitor PKI, was obtained from Addgene (#45066). pcDNA 3.1+ was from Invitrogen, USA. mCherry-3HA plasmid was generated in our laboratory.

Techniques: Expressing, Transfection, Plasmid Preparation, Isolation, Western Blot, Over Expression, Blocking Assay, Inhibition, Control, Two Tailed Test

Journal: Journal of cell science

Article Title: CREB regulates the expression of type 1 inositol 1,4,5-trisphosphate receptors.

doi: 10.1242/jcs.258875

Figure Lengend Snippet: Fig. 7. CREB is crucial in governing KRAP protein levels. (A) Representative western blot showing endogenous levels of KRAP protein induced with 10 µM forskolin (+) or not induced (−) in HEK-293 cells. (B) Quantitative analysis of three independent western blots as described under A. (C,E,G) HEK-293 cells were not transfected (0), or transiently transfected with increasing amounts (0.5, 1.0, 2.0 µg) of VP16-CREB (C), increasing amounts (0.25, 0.5, 1.0 µg) of KCREB (E) or increasing amounts (0.5, 1.0, 2.0 µg) of PKI (G) expression plasmid. Total protein amounts were isolated 36 h following transfection and analyzed by western blotting. Representative western blots are shown. (D,F,H) Quantitative analysis of three independent western blots for each experimental scenario as described under C, E and G. Endogenous levels of KRAP were increased upon overexpression of VP16-CREB (C), reduced when CREB was inhibited by overexpression of KCREB (E), and reduced when PKA was inhibited by overexpression of PKI (G). Statistical significance was determined by Student’s t-test (unpaired, two-tailed). *P<0.05, **P<0.01, ****P<0.0001.

Article Snippet: ITPR2 promoterreporter construct was kindly provided by Michael Nathanson (Yale University, New Haven, CT). pRL-TK plasmid was kindly provided by Cesare Orlandi (University of Rochester, Rochester, NY). pRSV-PKI-v2, expressing the PKA inhibitor PKI, was obtained from Addgene (#45066). pcDNA 3.1+ was from Invitrogen, USA. mCherry-3HA plasmid was generated in our laboratory.

Techniques: Western Blot, Transfection, Expressing, Plasmid Preparation, Isolation, Over Expression, Two Tailed Test

Journal: Cell reports

Article Title: EGFR-phosphorylated GDH1 harmonizes with RSK2 to drive CREB activation and tumor metastasis in EGFR-activated lung cancer

doi: 10.1016/j.celrep.2022.111827

Figure Lengend Snippet:

Article Snippet: Plasmid: pRSV-CaMKIV (1-313) , Sun et al. , Addgene plasmid: pRSV-CaMKIV(1-313); Cat#45063.

Techniques: Microarray, Recombinant, Purification, Membrane, SYBR Green Assay, Viability Assay, Phospho-proteomics, Kinase Assay, Reverse Transcription, Transcription Factor Assay, Enzyme-linked Immunosorbent Assay, Multiplex Assay, Extraction, Isolation, shRNA, Sequencing, Plasmid Preparation, Software