Journal: Oncotarget

Article Title: Long-term neurocognitive dysfunction in offspring via NGF/ ERK/CREB signaling pathway caused by ketamine exposure during the second trimester of pregnancy in rats

doi: 10.18632/oncotarget.16042

Figure Lengend Snippet: Ketamine administered during the second trimester perturbed NGF and down-regulated ERK, p-ERK, MSK, CREB, p-CREB, SYP, SYN, and PSD95 in the hippocampi of offspring. On postnatal day 30, after behavioral tests, brains were harvested for western blot analyses to determine the expression of NGF, ERK, p-ERK, MSK, CREB, p-CREB, SYP, SYN, and PSD95. (A–I ) Above are representative immunoblots for the expression levels of the corresponding proteins, and below are the histograms of the quantification of the corresponding proteins normalized to GAPDH. Data are expressed as mean percentage ± SEM of control mean values. * p < 0.05.

Article Snippet: The separated proteins were transferred onto nitrocellulose (NC) membranes and incubated with rabbit anti- GAPDH, NGF, ERK, p-ERK, CREB, p-CREB, MSK1, SYP, SYN, and PSD95 (1:600, EnoGene, Nanjing, China) overnight at a temperature of 4 °C.

Techniques: Western Blot, Expressing, Control

Journal: Oncotarget

Article Title: Long-term neurocognitive dysfunction in offspring via NGF/ ERK/CREB signaling pathway caused by ketamine exposure during the second trimester of pregnancy in rats

doi: 10.18632/oncotarget.16042

Figure Lengend Snippet: Ketamine administered during the second trimester down-regulated ERK, p-ERK, MSK, CREB, p-CREB, SYP, SYN, and PSD95 in PC12 cells during in vitro experiments. SCH772984 is the inhibitor of the ERK protein. Through the contrast between S group and C group, a very good effect of the inhibitor is revealed. (A) Representative immunoblots for expression levels of the corresponding proteins. (B–I ) Histograms that quantify the corresponding proteins normalized to GAPDH, in which the data are expressed as mean percentages ± SEM of control mean values. * p < 0.05

Article Snippet: The separated proteins were transferred onto nitrocellulose (NC) membranes and incubated with rabbit anti- GAPDH, NGF, ERK, p-ERK, CREB, p-CREB, MSK1, SYP, SYN, and PSD95 (1:600, EnoGene, Nanjing, China) overnight at a temperature of 4 °C.

Techniques: In Vitro, Western Blot, Expressing, Control

Journal: PLoS ONE

Article Title: Prostacyclin Analogue Beraprost Inhibits Cardiac Fibroblast Proliferation Depending on Prostacyclin Receptor Activation through a TGF β-Smad Signal Pathway

doi: 10.1371/journal.pone.0098483

Figure Lengend Snippet: Neonatal rat cardiac fibroblasts were pre-treated with beraprost (10 µM) for 4 h followed by Ang II (100 nM) stimulation for an additional 24 h. (A) Cell lysates were tested for phosphorylation of CREB at Ser133 and Ser142 by western blot. (B) Proteins from cytoplasm and nucleus respectively were tested for phosphorylation of CREB at Ser133 by western blot. (C) Nuclear proteins were immunoprecipitated with anti-CBP antibody. The precipitated proteins were analyzed with western blots using anti-Smad2 or Ser133-p-CREB antibodies. The total CBP detected with the anti-CBP antibody was used to normalize the signals. (D) Quantitative analysis of p-Smad2-CBP association. (E) Quantitative analysis of p-CREB-CBP association. Values are expressed as mean ± SEM. Cells treated with culture medium served as a vehicle control (con), * P <0.05, ** P <0.01, compared with con, # P <0.05, ## P <0.01, compared with only Ang II stimulated group. n = 4.

Article Snippet: After blocking at room temperature in 5% w/v non-fat milk with TBST buffer (Tris-HCl 10 mM, NaCl 120 mM, Tween-20 0.1%; pH 7.4) for 2 h, membranes were incubated overnight with the appropriate primary anti-collagen I, anti-collagen III, anti-TGF β, anti-Smad2, anti-p-Smad2, anti-CBP, anti-Ser142-p-CREB, anti-Ser133-p-CREB (1∶500, Bioworld Technology, St. Louis Park, MN), anti-tubulin, anti-Lamin B1(1∶1000, Santa Cruz Biotechnology, Santa Cruz, CA), anti-GAPDH (1∶6000, Sigma-Aldrich, St. Louis, MO) at 4°C and then incubated with horseradish peroxidase (HRP)-conjugated secondary antibody at room temperature for 2 h. Proteins were visualized by enhanced chemiluminescence substrate (ECL, Pierce, Rockford, IL).

Techniques: Western Blot, Immunoprecipitation