Journal: BioMed Research International

Article Title: Effects of Wenxin Keli on Cardiac Hypertrophy and Arrhythmia via Regulation of the Calcium/Calmodulin Dependent Kinase II Signaling Pathway

doi: 10.1155/2017/1569235

Figure Lengend Snippet: (a) The expression of CaMK II and related proteins in the CaMK II signal transduction pathway in the left ventricular areas after treatment for 3 and 9 weeks were analyzed by western blotting. (b) The expression of CaMK II ( n = 5). (c) The expression of p-CaMK II (Thr-286) ( n = 5). (d) The expression of PLB ( n = 5). (e) The expression of p-PLB (Thr-17) ( n = 5). (f) The expression of RyR2 ( n = 5). (g) The expression of p-RyR2 (Ser-2814) ( n = 5). ∗ P < 0.05 and ∗∗ P < 0.01 versus the Sham group. # P < 0.05 and ## P < 0.01 versus the TAC group.

Article Snippet: The antibodies to the following proteins were used: CaMK II delta (1 : 1000, Abcam Corporation.), phospho-CaMK II (1 : 1000, Cell Signaling Technology Inc.), PLB (G-18) (1 : 200, Santa Cruz Biotechnology Inc.), p-phospholamban-R (1 : 200, Santa Cruz Biotechnology Inc.), and ryanodine receptor 2 (1 : 1000, Millipore Corporation.).

Techniques: Expressing, Transduction, Western Blot

Journal: PLoS ONE

Article Title: Mu-Opioid Receptors Transiently Activate the Akt-nNOS Pathway to Produce Sustained Potentiation of PKC-Mediated NMDAR-CaMKII Signaling

doi: 10.1371/journal.pone.0011278

Figure Lengend Snippet: Upper panel : The influence of 10 nmol icv morphine on NMDAR activity was evaluated. The levels of NMDAR1 and of NMDAR2A subunits and activating phosphorylations at NR1 Ser890 (PKC site) and NR2A Tyr1325 (Src site) were monitored in the CD1 PAG synaptosomal membranes during the post-morphine intervals indicated. The presence of the Thr286 autophosphorylation of NMDAR-activated CaMKII was also determined. Administration of LNNA or naltrindole before morphine greatly reduced NR1, NR2A and CaMKII phosphorylation. Lower panel : Naltrindole decreased Akt/PKB activating phosphorylation. Please see for further details.

Article Snippet: Western blots were probed with affinity-purified IgG antibodies directed against: murine MOR (1∶1000 dilution) ; Gαi2, Gαz (1∶2000) ; RGS17(Z2) , ; NMDAR1 (1∶1000, Abcam ab1880); NMDAR1 phospho-Ser890 (Cell Signaling #3381); NMDAR2A (1∶1000, ab14596); NMDAR2A phospho-Y1325 (1∶300, ab16646); CaMKII (1∶3000; BD Transduction labs 611292); CaMKII phospho-Thr286 (1∶2000; Cell Signaling 3361); PKCγ (1∶1000; Abcam ab4145).

Techniques: Activity Assay

Journal: Chinese Medicine

Article Title: Chotosan ( Diaoteng San )-induced improvement of cognitive deficits in senescence-accelerated mouse (SAMP8) involves the amelioration of angiogenic/neurotrophic factors and neuroplasticity systems in the brain

doi: 10.1186/1749-8546-6-33

Figure Lengend Snippet: Effects of CTS on expression levels of p-NMDAR1, NMDAR1, p-CaMKII, CaMKIIα, p-CREB, CREB, and GAPDH in the cerebral cortex of SAMP8 with and without ischemic insult . Typical photos indicating the expression levels of each factor in the cerebral cortex of vehicle-treated SAMR1 control (lane 1), vehicle-treated sham-SAMP8 (lane 2), CTS (750 mg/kg/day)-treated sham-SAMP8 (lane 3), vehicle-treated T2VO-SAMP8 (lane 4), and CTS-treated T2VO-SAMP8 group (lane 5). After completing the behavioral studies, the animals were decapitated and proteins were extracted from the cerebral cortices in each animal group.

Article Snippet: They were then probed with anti-NMDAR1 rabbit polyclonal antibody (1:1000 dilution) and anti-phospho-NMDAR1 (p-NMDAR1) (Ser896) rabbit polyclonal antibody (1:1000 dilution), anti-CaMKIIα (A-1: sc-13141) mouse monoclonal antibody (1:1000 dilution), anti-phospho-CaMKII (p-CaMKII) (Thr286) rabbit polyclonal antibody (1:1000 dilution) (Cell Signaling Technology, USA), anti-CREB (48H2) rabbit monoclonal antibody (1:1000 dilution), anti-phospho-CREB (p-CREB) (Ser133) rabbit monoclonal antibody (1:1000 dilution), anti-BDNF (Tyr951) rabbit polyclonal antibody (1:500 dilution), anti-VEGF (A-20: sc-152) rabbit polyclonal antibody (1:1000 dilution) (Santa Cruz Biotechnology, USA), and anti-VEGFR2 (Ab-951) rabbit polyclonal antibody (1:1000 dilution) (Signalway Antibody, USA) and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mouse monoclonal antibody (1:2000 dilution) (Chemicon, USA) at 4°C for 24 hours.

Techniques: Expressing

Journal: Chinese Medicine

Article Title: Chotosan ( Diaoteng San )-induced improvement of cognitive deficits in senescence-accelerated mouse (SAMP8) involves the amelioration of angiogenic/neurotrophic factors and neuroplasticity systems in the brain

doi: 10.1186/1749-8546-6-33

Figure Lengend Snippet: Quantitative comparisons of CTS-induced changes in expression levels of p-NMDAR1, NMDAR1, p-CaMKII, CaMKIIα, p-CREB, CREB, and GAPDH in the cerebral cortex of SAMP8 with and without ischemic insult . The data are expressed as the percentage of the value obtained from naïve control SAMR1 mice. Each data column represents the mean ± SD obtained from 4-5 brain samples. # P < 0.05 or ## P < 0.01 vs. compared with vehicle-treated SAMR1 group (t-test). *** P < 0.001, ** P < 0.01 vs. respective vehicle-treated sham- or T2VO-SAMP8 group (two-way ANOVA).

Article Snippet: They were then probed with anti-NMDAR1 rabbit polyclonal antibody (1:1000 dilution) and anti-phospho-NMDAR1 (p-NMDAR1) (Ser896) rabbit polyclonal antibody (1:1000 dilution), anti-CaMKIIα (A-1: sc-13141) mouse monoclonal antibody (1:1000 dilution), anti-phospho-CaMKII (p-CaMKII) (Thr286) rabbit polyclonal antibody (1:1000 dilution) (Cell Signaling Technology, USA), anti-CREB (48H2) rabbit monoclonal antibody (1:1000 dilution), anti-phospho-CREB (p-CREB) (Ser133) rabbit monoclonal antibody (1:1000 dilution), anti-BDNF (Tyr951) rabbit polyclonal antibody (1:500 dilution), anti-VEGF (A-20: sc-152) rabbit polyclonal antibody (1:1000 dilution) (Santa Cruz Biotechnology, USA), and anti-VEGFR2 (Ab-951) rabbit polyclonal antibody (1:1000 dilution) (Signalway Antibody, USA) and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mouse monoclonal antibody (1:2000 dilution) (Chemicon, USA) at 4°C for 24 hours.

Techniques: Expressing

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: ANKRD22 Drives Rapid Proliferation of Lgr5 + Cells and Acts as a Promising Therapeutic Target in Gastric Mucosal Injury

doi: 10.1016/j.jcmgh.2021.06.020

Figure Lengend Snippet: ANKRD22 deletion suppresses the noncanonical Wnt-Ca 2+ pathway. ( A ) Effect of Ankrd22 knockout on the Wnt pathway in mouse gastric epithelium after mucosal injury detected by Western blot. ( B ) Effect of ANKRD22 on the Wnt pathway in cultured cells detected by Western blot. ( C ) Ankrd22 knockout reduced the intracellular Ca 2+ levels in mouse gastric epithelial cells detected by Fluo-4–based FCM. Ankrd22 +/+ and Ankrd22 -/- mice were subjected to subsequent detection at 24 hours after intragastric administration of EtOH/HCl or saline solution (Ctrl) (n = 4). ( D ) ANKRD22 increased the intracellular Ca 2+ levels in gastric cancer cells. ( E ) Mitochondrial colocalization of the exogenous-expressing ANKRD22 in the organoid-enriched gastric cancer cells detected by confocal microscopy. ( F and G ) Ankrd22 knockout reduced the mitochondrial Ca 2+ level in mouse gastric epithelium. Ankrd22 +/+ and Ankrd22 -/- mice were subjected to subsequent detection at 24 hours after intragastric administration of EtOH/HCl (n = 3). Confocal microscopy was used to compare the fluorescence intensity and colocalization with mitochondria. The fluorescence intensity was detected by Rhod-2–based FCM. ( H ) Inhibition of the Wnt-Ca 2+ pathway increased the canonical Wnt pathway activity in the gastric cancer cells detected by Western blot. ( I ) Inhibition of the Wnt-Ca 2+ pathway increased the number of organoid-enriched primary mouse gastric EPCs. ( H and I ) The cells were cultured for 24 hours in medium with or without (Ctrl) 10 μmol/L FK506 or KN93. ( J–L ) Ankrd22 knockout decreased the levels of diacylglycerol (DAG) phosphorylated-CaMKII, and NFAT in the mouse gastric epithelium after mucosal injury detected by Western blot (n = 3). ( M ) Expression of NFAT1 in the Ankrd22 +/+ and Ankrd22 -/- mouse gastric epithelium after mucosal injury detected by IHC staining. Normal mouse IgG was used as a negative control (Ctrl). Ankrd22 +/+ and Ankrd22 -/- mice were subjected to subsequent detection at 24 hours after intragastric administration of EtOH/HCl. The data were analyzed by the Student t test and are presented as means ± SD. ∗ P < .05 and ∗∗ P < .01. p-CaMKII, Phosphorylated calcium/calmodulin-dependent protein kinase type II.

Article Snippet: The following primary antibodies were used: β-tubulin (HuaBio), c-Myc (Cell Signaling Technology, Danvers, MA), AXIN2 (Cell Signaling Technology), NFAT1 (Proteintech, Wuhan, Hubei, China), NFAT2 (Cell Signaling Technology), NFAT3 (Abcam, Cambridge, UK), NFAT4 (GeneTex, Irvine, CA), diacylglycerol (LifeSpan BioSciences, Seattle, WA), phospho-CaMKII (Thr286; Cell Signaling Technology), and CaMKII (GeneTex).

Techniques: Knock-Out, Western Blot, Cell Culture, Expressing, Confocal Microscopy, Fluorescence, Inhibition, Activity Assay, Immunohistochemistry, Negative Control