factor human crh  (MedChemExpress)

Journal: Aging (Albany NY)

Article Title: Chronic stress induces Alzheimer’s disease-like pathologies through DNA damage-Chk1-CIP2A signaling

doi: 10.18632/aging.205862

Figure Lengend Snippet: Stress hormone activates DNA damage-Chk1-CIP2A pathway, results in tau hyperphosphorylation, Aβ overproduction and synaptic impairments in primary neurons. ( A – E ) Primary neurons were treated with 100 nM corticotropin-releasing factor (CRF) for 24 hours. ( A ) Representative immunoblots of γH2A.X, Chk1-pS317, Chk1-pS345, Chk1, CIP2A, Tau-pT231, Tau-pS396, Tau-pS404, Tau-5 (total tau), APP-pT668, APP and β-actin. ( B ) Quantification of the relative protein levels; non-phosphorylated proteins such as γH2A.X and CIP2A were normalized to the β-actin levels; phosphorylated Chk1-pS317, Chk1-pS345, Tau-pT231, Tau-pS396, Tau-pS404 and APP-pT668 were normalized to corresponding total Chk1, tau (Tau-5) and APP respectively. n = 3 or n = 6 per group. ( C ) The relative concentration of Aβ 40 and Aβ 42 in supernatant and cell lysate of primary neurons detected by ELISA. n = 3 or n = 4 per group. ( D ) Representative immunoblots of Synapsin I, PSD95, GluA1 and β-actin. ( E ) Quantification of the relative protein levels, which were normalized to the β-actin levels. n = 3 per group. Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Corticosterone (Cat# HY-B1618), Corticotropin-releasing factor (human) (Cat# HY-P0086) and GDC-0575 (Cat#HY-112167) were from MedChemExpress, USA.

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

Journal: Science Advances

Article Title: Hyperexcitation of ovBNST CRF neurons during stress contributes to female-biased expression of anxiety-like avoidance behaviors

doi: 10.1126/sciadv.adk7636

Figure Lengend Snippet: ( A ) Left: Representative image showing expression of CRFR1 in control mice. Right: Quantification of the expression of CRFR1 in female and male mice (** P < 0.01, unpaired t test). ( B ) Immunofluorescence staining of CRFR1 (green) and Cre (red) in the ovBNST of female (left) and male (right) CRF-Cre mice. Scale bars, 50 μm. ( C ) Quantification of CRFR1 neurons number between the ovBNST of female and male mice (unpaired t test). ( D ) Representative firing rate of ovBNST CRF neurons at base condition, after CRF perfusion and following R121919 perfusion. ( E ) Quantification of the firing rate of ovBNST CRF neurons at base condition, after CRF perfusion and following R121919 condition (*** P < 0.001, # P < 0.05, and ## P < 0.01, one-way ANOVA and post hoc test). ( F ) Experimental timeline. ( G ) Representative image (up) and quantification (down) of the expression of CRFR1 in control and CRFR1 knockdown female mice (* P < 0.05, unpaired t test). ( H ) Representative image showing expression of DIO-mCherry-shRNA3(CRFR1) in ovBNST. Scale bar, 200 μm. ( I and J ) Quantification of frequency to center zone and percentage time spent in the center zone of OFT (I), as well as quantification of frequency to open arm and percentage time spent in the open arm of EPM (J) in female mice (* P < 0.05 and ** P < 0.01, Student’s t test). ( K ) Representative image (up) and quantification (down) of the expression of CRFR1 in control and CRFR1 overexpression female mice (* P < 0.05, unpaired t test). ( L ) Representative image showing expression of DIO-crfr1-EGFP in ovBNST. Scale bar, 200 μm. ( M and N ) Quantification of frequency to center zone and percentage time spent in the center zone of OFT (M), as well as quantification of frequency to open arm and percentage time spent in the open arm of EPM (N) in male mice (* P < 0.05 and *** P < 0.001, Student’s t test). GAPDH, glyceraldehyde phosphate dehydrogenase. KD, knock down; KO, knock out; OE, over expression.

Article Snippet: For drug administration, 15 min before the forced swimming stress, CRF (1 μg in 0.3 μl per site, MedChemExpress) or R121919 (1 μg in 0.3 μl per site, MedChemExpress) was infused through the internal cannula inserted in the guide cannula, while control animals received saline solution.

Techniques: Expressing, Control, Immunofluorescence, Staining, Knockdown, Over Expression, Knock-Out

Journal: Journal of Cellular and Molecular Medicine

Article Title: Targeting the TR4 nuclear receptor with antagonist bexarotene can suppress the proopiomelanocortin signalling in AtT‐20 cells

doi: 10.1111/jcmm.16074

Figure Lengend Snippet: Bexarotene treatment reduces POMC transcription and ACTH secretion in AtT‐20 cells. A, Changes in POMC and TR4 mRNA after treatment with 0, 5 and 10 µmol/L bexarotene for 12 h were assessed by qPCR; B, Protein levels of POMC and TR4 after treatment with 0, 5 and 10 µmol/L bexarotene for 24 h were assessed by Western blot analysis. C, Cells were transfected with a POMC promoter‐luciferase reporter and pRL‐TK(Δ238); after 18 h, cells were treated with 0, 5 or 10 µmol/L bexarotene for 24 h, and luciferase activities were measured and presented as the fold change compared the control; D, AtT‐20 cells were treated for 24 h with 0, 5 or 10 µmol/L bexarotene, 40 µmol/L MEK‐162, 100 nmol/L CRH, 100 nmol/L DEX, or their combinations as indicated. ACTH levels were measured in the supernatants from AtT‐20 cells through ECLIA. Results are expressed as the percentages of each control. Data are the means * SEM of three independent experiments. * P ˂ .05, ** P ˂ .01 except in Figure * P ˂ .01, compared with the basal values, ** P ˂ .01 compared with CRH stimulation, *** P ˂ .01 compared with DEX suppression. One‐way ANOVA and two‐way ANOVA were used to test differences for statistical significance

Article Snippet: Corticotropin‐releasing factor human (CRH) was obtained from MCE and stored at −80°C.

Techniques: Western Blot, Transfection, Luciferase, Control