Journal: Frontiers in Pharmacology

Article Title: CyPPA, a Positive SK3/SK2 Modulator, Reduces Activity of Dopaminergic Neurons, Inhibits Dopamine Release, and Counteracts Hyperdopaminergic Behaviors Induced by Methylphenidate ¹

doi: 10.3389/fphar.2012.00011

Figure Lengend Snippet: CyPPA inhibits spontaneous and evoked release of dopamine form TH/SK3 positive cultured rat midbrain neurons . (A) Western blot detection of SK3 in rat midbrain cultures. Extracts of HEK293 cells transiently transfected with rSK1, rSK2, and rSK3 and adult rat brain membranes (RBM) as well as lysates from cultured midbrain neurons were separated on 4–15% SDS-PAGE and immunoblotted with anti-SK3 antibody. A band of approximately 70 kDa was detected in midbrain cultures, RBM lysates, and HEK293 cells transfected with rSK3. This band was absent in lanes loaded with HEK293 cells expressing rSK1 and rSK2. (B) Immunodetection of SK3 subunits in rat midbrain cultures. Cultured midbrain neurons (8 days in vitro ) were double-labeled for SK3 and tyrosine hydroxylase (TH). In TH-positive dopaminergic neurons, the SK3 subunit displayed a primarily somatic–dendritic localization whereas little immunoreactivity was associated with axons (arrow). Scale bar 20 μm. (C) Basal release and stimulated release with 25 mM KCl or inhibited release with 30 μM Cd 2+ and with 1 μM TTX. (D) Basal release and stimulated release with 3 μM haloperidol and inhibited release with 10 μM quinpirole. Blue bars represent inhibited release with increasing concentrations of CyPPA (* P < 0.05, ** P < 0.01). The data presented are representative of three independent experiments.

Article Snippet: Neurobasal media and B-27 were obtained from Invitrogen (Nærum, Denmark) and 3 H-DA was acquired from PerkinElmer Life and Analytical Sciences (Boston, MA, USA) The primary antibodies used were a rabbit polyclonal anti-SK3 antibody (1:100, APC-025) from Alomone Labs (Jerusalem, Israel) and a monoclonal mouse anti-tyrosine hydroxylase antibody (1:150, clone LNC1) from Millipore (Copenhagen, Denmark).

Techniques: Cell Culture, Western Blot, Transfection, SDS Page, Expressing, Immunodetection, In Vitro, Labeling

Journal: Bioactive Materials

Article Title: A continuous adhesion-enhanced osteogenic pathway in artificial scaffold drives cellular infiltration and condensed mineralization for rapid bone regeneration

doi: 10.1016/j.bioactmat.2026.02.026

Figure Lengend Snippet: IHC and IF staining of functional proteins. (a) H&E and Masson's staining of defect area after implantation of CPH/rGO-3/0 and CPH/rGO-3/0.6 scaffolds for one month. (b) OCN IHC and IF staining of CPH/rGO-3/0 and CPH/rGO-3/0.6 after implantation for one month and the corresponding OD value and fluorescence intensity statistics. (c) IF staining of FREM1 and ANK3 in scaffolds after implanting for one months in rats and corresponding fluorescence intensity statistics. (d) IF staining of KCNN3 and KCNH1 in scaffolds after implanting for one months in rats and corresponding fluorescence intensity statistics. n = 20 per group. Data are expressed as mean ± SD. ∗ for p < 0.05; ∗∗ for p < 0.01; ∗∗∗ for p < 0.001.

Article Snippet: Hematoxylin-Eosin (Solarbio, Beijing, China) Stain Kit, Masson's Trichrome Stain Kit (Solarbio, Beijing, China), OCN monoclonal antibody (ThermoFisher Scientific, USA) FREM1 and KCNN3 antibody (Bioss Antibodies, Beijing, China), ANK3 antibody (Proteintech, Wuhan, China), KCNH1 (Servicecbio, Wuhan, China) were used to stain the slice of specimens.

Techniques: Staining, Functional Assay, Fluorescence