Journal:

Article Title: Activation of m-Calpain (Calpain II) by Epidermal Growth Factor Is Limited by Protein Kinase A Phosphorylation of m-Calpain

doi: 10.1128/MCB.22.8.2716-2727.2002

Figure Lengend Snippet: Effects of CPT-cAMPS and PKA inhibitors on EGF-induced cell migration. NR6WT cells were grown to confluence and made quiescent for 24 h in MEMα with 0.5% dialyzed fetal calf serum before treatment or not with EGF (10 nM), CPT-cAMPS (1 μM), Rp-8Br-cAMPS and/or Rp-8Br-cGMPS (5 μM) (A), and H-89 (at indicated concentrations) (B). The datum for nontreated control cells is labeled as nTx. Basal and EGF-induced cell migrations were assessed as the ability of the cells to move into an acellular area after 24 h of EGF treatment. The data are shown as the ratios to the 10 nM EGF-induced cell migration activity. The data are the means ± SEM of at least three independent studies, each performed in triplicate. Statistical analysis was performed by Student's t test. n.s., not significant.

Article Snippet: Cells were treated or not with CPT-cAMPS (1 μM) and/or Rp-8Br-cAMPS (5 μM) and incubated for 30 min in the presence of 30 μM BOC-LM-CMAC (Molecular Probes, Eugene, Oreg.).

Techniques: Migration, Labeling, Activity Assay

Journal:

Article Title: Activation of m-Calpain (Calpain II) by Epidermal Growth Factor Is Limited by Protein Kinase A Phosphorylation of m-Calpain

doi: 10.1128/MCB.22.8.2716-2727.2002

Figure Lengend Snippet: EGF-induced calpain activation. NR6WT cells were plated on tissue culture chamber slides (Nunc) and made quiescent for 24 h in MEMα with 0.5% dialyzed fetal calf serum. Cells were treated or not with CPT-cAMPS (1 μM), Rp-8Br-cAMPS (5 μM), and/or Rp-8Br-cGMPS (5 μM) 30 min prior to EGF (10 nM) treatment in the presence of BOC-LM-CMAC (Molecular Probes). Then cells were treated or not with EGF (10 nM) for 5 min. Calpain activation was assessed by fluorescence microscopy. The fluorescence indicates calpain activity. The panel for nontreated control cells is labeled as nTx. The pictures shown are representative of n = 9.

Article Snippet: Cells were treated or not with CPT-cAMPS (1 μM) and/or Rp-8Br-cAMPS (5 μM) and incubated for 30 min in the presence of 30 μM BOC-LM-CMAC (Molecular Probes, Eugene, Oreg.).

Techniques: Activation Assay, Fluorescence, Microscopy, Activity Assay, Labeling

Journal: American Journal of Physiology - Renal Physiology

Article Title: Role of cAMP/PKA signaling cascade in vasopressin-induced trafficking of TRPC3 channels in principal cells of the collecting duct

doi: 10.1152/ajprenal.00586.2009

Figure Lengend Snippet: Effect of PKA inhibitors Rp-cAMPS and myristoylated inhibitor of PKA (m-PKI) on TRPC3 translocation. A: protocol was identical to that described in the legend to Fig 3. IMCD-3 cells were pretreated with Rp-cAMPs (10 μM) or m-PKI (1 μM) for 2 h before addition of AVP or forskolin to the bath solution. The cell monolayers were fixed and labeled with primary antibodies against TRPC3 (green) and AQP2 (red). The figure shows representative merged confocal images taken 30 min after addition of AVP or forskolin as indicated. B: surface biotinylation assays were performed as described in the legend to Fig 7 in control IMCD-3 cells or in cells pretreated with the PKA inhibitors as indicated above each lane. C: bands from 3 independent experiments were quantified by densitometry. The intensity of the TRPC3 bands from the avidin pulldowns were normalized to the value obtained for the corresponding TRPC3 IP bands under each condition.

Article Snippet: The myristoylated inhibitor of PKA (m-PKI) and Rp-cAMPS were from Biomol.

Techniques: Translocation Assay, Labeling, Avidin-Biotin Assay

Journal: The European journal of neuroscience

Article Title: Transient viral-mediated overexpression of α-calcium/calmodulin-dependent protein kinase II in the nucleus accumbens shell leads to long-lasting functional upregulation of α-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate receptors: dopamine type-1 receptor and protein kinase A dependence

doi: 10.1111/j.1460-9568.2010.07155.x

Figure Lengend Snippet: Location of microinjection cannula tips in the NAcc shell for all rats included in the data analyses. Line drawings of coronal sections (adapted from Paxinos and Watson, 1997) show the location of the microinjection cannula tips in the NAcc shell for rats tested with no inhibitor (A), with SCH23390 (B) or with Rp-cAMPS (C). Numbers to the right indicate mm from bregma. Symbols denote group affiliation: filled circles, αCaMKII-AMPA; open circles, control-AMPA; filled squares, αCaMKII-saline; open ssquares, control-saline.

Article Snippet: Rats in separate groups were administered saline (0.5 µL/side) or the AMPA receptor agonist (±) AMPA hydrobromide (0.8 nmol/0.5 µL/side; Sigma-Aldrich Inc.) infused either alone or with the D1 receptor antagonist SCH23390 HCl (0.8 nmol/0.5 µL/side; Tocris Bioscience, Ellisville, MO, USA) or the PKA inhibitor Rp-cAMPS (80 nmol/1 µL/side; Axxora LLC, San Diego, CA, USA).

Techniques:

Journal: The European journal of neuroscience

Article Title: Transient viral-mediated overexpression of α-calcium/calmodulin-dependent protein kinase II in the nucleus accumbens shell leads to long-lasting functional upregulation of α-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate receptors: dopamine type-1 receptor and protein kinase A dependence

doi: 10.1111/j.1460-9568.2010.07155.x

Figure Lengend Snippet: Enhanced locomotor responding to NAcc shell AMPA at 20 days post-infection requires D1 receptor and PKA activation. Data in (A–F) are shown as group mean (±SEM) locomotor counts obtained before and after the challenge injection (arrows). (A and B) The locomotor response to NAcc shell AMPA was significantly enhanced in αCaMKII compared with control group rats. Little locomotor activity was observed in either group following NAcc shell saline. Coadministration of either SCH23390 (C and D) or Rp-cAMPS (E and F) with AMPA into the NAcc shell blocked the enhanced locomotor response in αCaMKII compared with control group rats but spared the locomotor effects of AMPA in both groups compared with NAcc shell saline. Neither SCH23390 nor Rp-cAMPS produced effects that differed significantly from saline when administered alone. (G) Summary of post-challenge injection results illustrated as group mean (+SEM) 2 h total locomotor counts. *P < 0.001, AMPA vs. saline; †P < 0.01, αCaMKII vs. control; revealed by post-hoc Scheffé comparisons following anova.

Article Snippet: Rats in separate groups were administered saline (0.5 µL/side) or the AMPA receptor agonist (±) AMPA hydrobromide (0.8 nmol/0.5 µL/side; Sigma-Aldrich Inc.) infused either alone or with the D1 receptor antagonist SCH23390 HCl (0.8 nmol/0.5 µL/side; Tocris Bioscience, Ellisville, MO, USA) or the PKA inhibitor Rp-cAMPS (80 nmol/1 µL/side; Axxora LLC, San Diego, CA, USA).

Techniques: Infection, Activation Assay, Injection, Activity Assay, Produced