Journal: Scientific Reports

Article Title: Treatment of beta amyloid 1–42 (Aβ 1–42 )-induced basal forebrain cholinergic damage by a non-classical estrogen signaling activator in vivo

doi: 10.1038/srep21101

Figure Lengend Snippet: Graphs show the percentage of ERK1/2 ( A1 ) and CREB ( B1 ) phosphorylation in ChAT neurons within the NBM 30 min after estren administration. Photomicrographs demonstrate dual-label immunofluorescence of pERK1/2 ( A2 – A4 ) or pCREB ( B2 – B4 ) immunoreactivity (red) in ChAT neurons (green). Bar graphs showing the percentage of ChAT neurons expressing pERK1/2 immunoreactivity in the NBM 30 min after incubation in vitro in ACSF containing vehicle (open bars) or estren (closed bars) with or without 0.5 μM TTX and amino acid receptor blocker cocktail (10 μM CNQX, 20 μM AP5, 50 μM picrotoxin) ( C ). The effect of estren treatment on cholinergic cells and fibers after Aβ 1–42 lesion in neuron-specific ERα knock out mice ( D ). Scale bars, 20 μm (A2-A4) and 40 μm ( B2 – B4 ). Histograms show mean ± SEM (n = 5–6). *P < 0.05; **P < 0.01; ***P < 0.001 (ANOVA with post hoc Bonferroni test).

Article Snippet: Briefly, brain sections were incubated with one of the primary antibodies recognizing pERK1/2, ERK1/2, pCREB or CREB (pERK1/2, 1:500, ERK1/2, 1:500, pCREB, 1:100, CREB, 1:1000; Cell Signaling Technologies; Aβ 1–42 , 1:500, Thermo Fisher Scientific) followed by incubation with chicken anti-rabbit Alexa Fluor 647 secondary antibody (1:500; Life Technologies).

Techniques: Immunofluorescence, Expressing, Incubation, In Vitro, Knock-Out

Journal: bioRxiv

Article Title: β2-Adrenergic Biased Agonist Nebivolol Inhibits the Development of Th17 and the Response of Memory Th17 Cells in an NF-κB-Dependent Manner

doi: 10.1101/2024.09.08.611829

Figure Lengend Snippet: Blasted memory Th cells activated with antiCD3/antiCD28/antiCD2 for 15 minutes in conditions of non-activated cells, activated cells, activated cells plus nebivolol, terbutaline A) Representative western blot data of equal amounts of protein from the cell lysates was shown for p-CREB and α-tubulin as a loading control. B) Band intensity was quantified and shown corrected to the loading control as a ratio, with data pooled from three independent biological experiments. C) Memory Th cells were activated for five days, with nebivolol plus or minus H89. A representative IL-17A ELISA data for the effect of PKA in nebivolol’s signaling pathway by using H89 as a PKA inhibitor. ANOVA was followed by Tukey’s multiple comparisons tests.

Article Snippet: Antibodies were against human Phospho-ser529 NF-κB p65 (1/500, clone A21012B Biolegend), NF-κB p65 Antibody (1/1000, clone 14G10A21, Biolegend), phospho-Ser133 CREB1 (1/700, rabbit polyclonal, Cusabio, Cedarlane), phospho-Ser412-β-arrestin1 (1/1000, clone mAb #2416 Cell Signaling Tech., Danvers, USA), β2AR (1/500, clone 4A6C9, Novusbio, Cedarlane), GAPDH (1/15000, 10B4E3, MA000071M1m, Cusabio, Cedarlane) and α□tubulin (1/1000, Santa Cruz Biotechnology Inc., USA).

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

Journal: Journal of neurochemistry

Article Title: Serotonergic Psychedelics Rapidly Modulate Evoked Glutamate Release in Cultured Cortical Neurons.

doi: 10.1111/jnc.70020

Figure Lengend Snippet: FIGURE 1 | Serotonergic psychedelics increase nuclear localization of phosphorylated CREB after 24 h treatment. Representative images of im- munofluorescent detection of pCREB Ser133 (pseudocoloured, range shown in the rectangle below) and dendritic marker MAP2 (red) overlaid with DAPI nuclear staining (blue) in mature cortical cultures treated with CTRL or drugs for 30 min (a), 2 h (c) or 24 h (e). Scale bar is 15 μm. Quantification of nuclear immunofluorescence (IF) of pCREB after treatment with CTRL or drugs for 30 min (b), 2 h (d) or 24 h (f), number of visual fields (each consisting of 10–15 analysed nuclei) from 2 to 3 independent experiments in brackets. Data did not pass Shapiro–Wilk test for normality, and were assessed by Kruskal–Wallis test (30 min: H(3,92) = 7.020, p = 0.071; 2 h: H(3,108) = 1.003, p = 0.801; 24 h: H(3,128) = 40.26, p < 0.001) and Dunn's multiple comparisons test of CTRL versus each drug (24 h: p < 0.001 for all comparisons).

Article Snippet: Antibodies used for immunocytochemistry experiments: phospho- CREB (Ser133; 1:700, Cell Signaling Technology, RRID:AB_2561044) and MAP2 (1:500, Sigma- Aldrich, RRID:AB_477193).

Techniques: Marker, Staining, Immunofluorescence