Journal: Iranian Journal of Basic Medical Sciences

Article Title: Adrenomedullin increases cAMP accumulation and BDNF expression in rat DRG and spinal motor neurons

doi: 10.22038/ijbms.2021.54796.12289

Figure Lengend Snippet: Level of phosphorylated-cAMP response element-binding protein (p-CREB) was increased in dorsal root ganglion (DRG) (A) and spinal motor neurons (B) following treatment with adrenomedullin (AM;25 nM). ELISA was used to measure the level of p-CREB (Ser133) in both DRG (A) and spinal motor neurons (B) following exposure to AM and forskolin. Pretreatment with AM22-52 reversed the AM-induced p-CREB elevation in both DRG (A) and spinal motor neurons (B). The data were analyzed using one way-ANOVA, followed by Dunnett's post hoc tests. Data are presented as mean±SEM, and at least three independent experiments were performed

Article Snippet: The p-CREB (Ser133) level was assayed in the cell lysate by DuoSet IC Phospho-CREB (S133) ELISA kit (R&D Systems).

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of Leukocyte Biology

Article Title: VIP plasma levels associate with survival in severe COVID‐19 patients, correlating with protective effects in SARS‐CoV‐2‐infected cells

doi: 10.1002/JLB.5COVA1121-626R

Figure Lengend Snippet: VIP and PACAP regulate the activation of transcription factors in SARS‐CoV‐2‐infected monocytes. Monocytes were treated (overnight) or not with to VIP or PACAP (10 nM), culture medium was removed and then cells were infected with SARS‐CoV‐2 for 1 h, as described in Material and Methods . After infection, viral input was removed, and cells were washed and then reexposed to the neuropeptides. After 24 h, cells were lysed and the ratios between phosphoNF‐kBp65 and total NF‐kBp65 (A), phosphoCREB and total CREB (B), active SREBP‐1 and β‐actin (C), and active SREBP‐2 and β‐actin (D) were quantified by ELISA (A and B) or by Western blot (C and D) in the cell lysates. Data represent means ± sd . */#, p ≤ 0.05; **/##, p ≤ 0.01; ***/###, p ≤ 0.001; ns, not significant; (A, B, C, and D) n = 4. For A and B, each dot represents an independent assay with 2 replicates

Article Snippet: Evaluation of NF‐kBp65 and CREB activation was performed in infected or uninfected monocytes using NF‐kBp65 (Total/Phospho) InstantOneTM and CREB (Total/Phospho) Multispecies InstantOneTM ELISA Kits (Thermo Fisher), according to manufacturer's instructions.

Techniques: Activation Assay, Infection, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: bioRxiv

Article Title: VIP plasma levels associate with survival in severe COVID-19 patients, correlating with protective effects in SARS-CoV-2-infected cells

doi: 10.1101/2020.07.25.220806

Figure Lengend Snippet: Monocytes were treated (overnight) or not with to VIP or PACAP (10 nM), culture medium was removed and then cells were infected with SARS-CoV-2. After infection, viral input was removed, and cells were washed and then re-exposed to the neuropeptides. After 24 hours, cells were lysed and the ratios between phosphoNF-kBp65 and total NF-kBp65 ( A ), phosphoCREB and total CREB ( B ), active SREBP-1 and β-actin ( C ), and active SREBP-2 and β-actin ( D ) were quantified by ELISA ( A, B ) or by western blot ( C, D ) in the cell lysates. Data represent means ± SD. * p ≤ .05; ** p ≤ .01; *** p ≤ .001; (A, B) n=3; (C, D) n=4. Each dot represents an independent assay with three replicates.

Article Snippet: Evaluation of NF-kBp65 and CREB activation was performed in infected or uninfected monocytes using NF-kB p65 (Total/Phospho) InstantOne™ and CREB (Total/Phospho) Multispecies InstantOne™ ELISA Kits (Thermo Fisher), according to manufacturer’s instructions.

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Scientific Reports

Article Title: Fermented Sipjeondaebo-tang Alleviates Memory Deficits and Loss of Hippocampal Neurogenesis in Scopolamine-induced Amnesia in Mice

doi: 10.1038/srep22405

Figure Lengend Snippet: ( A ) Levels of hippocampal BDNF from each mouse group were determined quantitatively using the ELISA assay. Data are presented as the mean ± SEM ( n = 6–8 mice/group). **p < 0.01, compared with CON-treated group; # p < 0.05, compared with SCO-treated group. ( B ) The representative images show DAB immunostaining for pCREB in the hippocampal DG for each 14 day drug treatment. The scale bar = 100 μm. The graph shows the number of pCREB-positive cells in the DG. Data are presented as the mean ± SEM (n = 5 mice/group). **p < 0.01, compared with CON-treated group; ## p < 0.01, compared with SCO-treated group. ( C ) The effects of SJ or FSJ on pCREB expression in the hippocampus of mice administered SCO are shown. A representative immunoblot from one of three independent experiments shows the results for the antibodies against pCREB, CREB, and β-actin. The lower graph shows the quantification of pCREB expressed as the ratio of pCREB/CREB. Data are presented as the mean ± SEM ( n = 6–8 mice/group). **p < 0.01, compared with CON-treated group; # p < 0.05, ## p < 0.01, compared with SCO-treated group. ( D ) The effects of SJ and FSJ on pAkt expression in the hippocampus in mice treated with SCO are shown. A representative immunoblot from one of three independent experiments shows the results for the antibodies against pAkt, Akt, and β-actin. The lower graph shows the quantification of pAkt expressed as the ratio of pAkt/Akt. Data are presented as the mean ± SEM ( n = 6–8 mice/group). **p < 0.01, compared with CON-treated group; ## p < 0.01, compared with SCO-treated group.

Article Snippet: Anti-phosphorylated (p) Akt (ser 473), total Akt, pCREB (ser 133), total CREB were purchased from Cell Signaling Technology.

Techniques: Enzyme-linked Immunosorbent Assay, Immunostaining, Expressing, Western Blot