Journal: PLoS ONE

Article Title: Desipramine Protects Neuronal Cell Death and Induces Heme Oxygenase-1 Expression in Mes23.5 Dopaminergic Neurons

doi: 10.1371/journal.pone.0050138

Figure Lengend Snippet: Cells were pretreated with various of MAP kinase inhibitors SB203580 (10 µM), PD98059 (20 µM) or SP600125 (10 µM) for 30 min followed by stimulation with desipramine (20 µM; A) or fluoxetine (20 µM; D) for another 24 h. Whole cell lysates were subjected to Western blot for detection of HO-1 expression. Cells were incubated with desipramine or fluoxetine for the indicated time periods, and cell lysates were subjected to immunoblots with antibodies against phospho-JNK (B and C) and phospho-ERK (D and E). Results are the representatives of three or four independent experiments.

Article Snippet: SP600125 was obtained from Tocris Bioscience (Ellisville, MO).

Techniques: Western Blot, Expressing, Incubation

Journal: PLoS ONE

Article Title: Desipramine Protects Neuronal Cell Death and Induces Heme Oxygenase-1 Expression in Mes23.5 Dopaminergic Neurons

doi: 10.1371/journal.pone.0050138

Figure Lengend Snippet: (A) Cells were treated with desipramine (20 µM) for indicated time periods (60 or 120 min) and nuclear extracts were collected, and the binding activity of Nrf2 to Nrf2-DNA binding element was examined by EMSA analysis. The DNA binding activity of Nrf2 is significantly different between desipramine treatment group and control group (one-way ANOVA followed by Bonferroni’s post hoc test). Cells were pretreated with PD98059 or SP600125 with desipramine (20 µM), and nuclear extracts were examined by EMSA analysis. Lane 1 was loaded without nuclear extracts (probe only). Results are expressed as the means ± S.E.M. from three independent experiments. *, p <0.05 as compared with the vehicle control group. #, p<0.05 as compared with the desipramine treatment group. (B) Cells were transfected with Control siRNA (100 nM) or Nrf2 siRNA (50 and 100 nM) for 24 h followed by stimulation with desipramine (20 µM) for another 24 h, and the protein levels of Nrf2 and HO-1 were determined by Western blot. The HO-1 expression is significantly different between Nrf2 siRNA group and control siRNA group (one-way ANOVA followed by Bonferroni’s post hoc test). Results are expressed as the means ± S.E.M. from three independent experiments. *, p <0.05 as compared with the control group. #, p<0.05 as compared with the desipramine treatment alone group.

Article Snippet: SP600125 was obtained from Tocris Bioscience (Ellisville, MO).

Techniques: Binding Assay, Activity Assay, Control, Transfection, Western Blot, Expressing

Journal: Critical Care Explorations

Article Title: Surfactant Protein D Influences Mortality During Abdominal Sepsis by Facilitating Escherichia coli Colonization in the Gut

doi: 10.1097/CCE.0000000000000699

Figure Lengend Snippet: Surfactant protein D (SPD) is synthesized by the gallbladder and promotes colonization of both the cecum and colon with Escherichia coli . A , SPD +/+ gut organs were harvested at baseline or post-cecal ligation and puncture (CLP) with Western blots performed for SPD or β -actin (loading control). Blots represent pooled samples, n = 2/group. Representative gel shown from three experiments. Lungs from SPD −/− and SPD +/+ mice were used as negative and positive controls, respectively. B , Gallbladder was isolated from SPD +/+ mice after CLP or sham surgery and from SPD −/− mice after CLP with Western blots performed for SPD or glyceraldehyde 3-phosphate dehydrogenase antibody (GAPDH; loading control). Blots represent pooled samples, n = 5–7/group. Lungs from SPD −/− mice and SPD +/+ were used as negative and positive controls, respectively. C , SPD −/− mice ( n = 9) were gavaged with recombinant surfactant protein D (rSPD), followed by gavage with green fluorescent protein (GFP)-labeled E. coli , and compared with SPD −/− mice gavaged only with GFP-labeled E. coli ( n = 9). After 24 hr, cecum and colon were harvested. GFP-labeled E. coli were then detected by culture (Mann-Whitney * p < 0.05, ** p < 0.01). CFU = colony forming units.

Article Snippet: SPD +/+ and SPD −/− mice were gavaged with 10 μL/g of ampicillin-resistant GFP-labeled E. coli (ATCC 25922GFP, 3.74 × 10 8 colony forming units/mL) with or without recombinant human surfactant protein D (rSPD) (Cat 1920-SP-050, R&D systems).

Techniques: Synthesized, Ligation, Western Blot, Control, Isolation, Recombinant, Labeling, MANN-WHITNEY