Journal: Scientific Reports

Article Title: The Na +/ H + -Exchanger NHE1 Regulates Extra- and Intracellular pH and Nimodipine-sensitive [Ca 2+ ] i in the Suprachiasmatic Nucleus

doi: 10.1038/s41598-019-42872-w

Figure Lengend Snippet: NHE1 colocalisation with CaV1.2 ( A ) and NCX1 ( B ), as well as colocalisation of NHE1/CaV1.2 with SERT ( C ), NPY ( D ), and Bassoon ( E ). ( A , B ) High magnification images showing high levels of NHE1 colocalisation with CaV1.2 ( A ), but not NCX1 ( B ). ( C – E ) High magnification images showing moderate to high levels of NHE1/CaV1.2 colocalisation with SERT ( C ) and Bassoon ( E ), but not NPY ( D ). Scale bar: 10 µm.

Article Snippet: For immunofluorescence staining, sections (20 µm) were washed for 20–30 min in PBS and then incubated overnight at 4 °C in PBS containing 2% serum, 0.3% Triton X-100, and primary antibodies against NHE1 (rabbit anti-NHE1; 1:100; ab67314, RRID:AB_1141782; Abcam, Cambridge, MA, USA), neurophysin II (NP2) (goat anti-NP2; 1:500; sc-27093, RRID:AB_2061964; Santa Cruz, CA, USA), vasoactive intestinal peptide (VIP) (guinea pig anti-VIP; 1:500; T-5030, RRID:AB_518690; Peninsula Laboratories, San Carlos, CA, USA), gastrin-releasing peptide (GRP) (goat anti-GRP; 1:100; sc-7788, RRID:AB_2232721; Santa Cruz, CA, USA), vesicular glutamate transporter type 2 (vGluT2) (guinea pig anti-vGluT2; 1:300; AB2251, RRID:AB_1587626; Millipore, Temecula, CA, USA), serotonin transporter (SERT) (mouse anti-SERT; 1:200; MAB1564, RRID:AB_94220; Millipore, Temecula, CA, USA), neuropeptide Y (NPY) (goat anti-NPY; 1:300; NBP1-46535, RRID:AB_10009813; Novus, Littleton, CO, USA), CaV1.2 (guinea pig anti-CaV1.2; 1:100; AGP-001; RRID:AB_11219156; Alomone Labs, Jerusalem, Israel), NCX1 (mouse anti-NCX1, against epitope between amino acid 371 and 525 on intracellular side of plasma membrane; 1:100; AB2869, RRID:AB_2191134; Abcam, MA, USA), and Bassoon (mouse anti-Bassoon; 1:200; ADI-VAM-PS003-D, RRID:AB_2038857; Enzo Life Sciences, Farmingdale, NY, USA).

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Article Title: Cathepsin-L, a Key Molecule in the Pathogenesis of Drug-Induced and I-Cell Disease-Mediated Gingival Overgrowth

doi:

Figure Lengend Snippet: Latent and active cathepsin activity of cells cultured with or without nifedipine. Enzyme activities of active forms of cathepsin-(B+L) (A), active form of cathepsin-B (B), total forms of cathepsin-(B+L) (C), and total form of cathepsin-B (D) were measured in gingival fibroblasts cultured with or without 100 ng/ml of nifedipine. Gingival fibroblasts cultured with or without nifedipine were subjected to measurement of cathepsin activities as described in Materials and Methods. Values are representative of the results obtained from gingival fibroblasts isolated from normal gingival tissues and nifedipine-induced gingival overgrowth, since gingival fibroblasts obtained from nifedipine-induced gingival overgrowth and from normal tissues were similarly affected by nifedipine. Values are expressed as the % activity against that of controls (day 0). Both active and total forms of cathepsin-(B+L) activity were significantly suppressed by nifedipine, while that of cathepsin-B was not influenced.

Article Snippet: Reagents The blocker of L-type Ca (2+) channels, nifedipine, was from Bayer AG Pharma Research Center (Wuppertal, Germany).

Techniques: Activity Assay, Cell Culture, Isolation

Journal:

Article Title: Cathepsin-L, a Key Molecule in the Pathogenesis of Drug-Induced and I-Cell Disease-Mediated Gingival Overgrowth

doi:

Figure Lengend Snippet: mRNA expression of various lysosomal enzymes cultured with or without nifedipine in gingival fibroblasts. The expressions of mRNA encoding various lysosomal enzymes were examined in gingival fibroblasts cultured with (nifedipine) or without (−) 100 ng/ml of nifedipine for the indicated time periods. Total RNA was isolated and was subjected to RT-PCR analysis as described in Materials and Methods. Values are representative of the results obtained from several gingival fibroblasts from different donors, since all cells tested showed similar results. Expression of cathepsin-L mRNA was markedly suppressed in gingival fibroblasts after 1 day of incubation with nifedipine, while that of other lysosomal enzymes was not apparently influenced. HeLa cells served as the positive control.

Article Snippet: Reagents The blocker of L-type Ca (2+) channels, nifedipine, was from Bayer AG Pharma Research Center (Wuppertal, Germany).

Techniques: Expressing, Cell Culture, Isolation, Reverse Transcription Polymerase Chain Reaction, Incubation, Positive Control

Journal:

Article Title: Cathepsin-L, a Key Molecule in the Pathogenesis of Drug-Induced and I-Cell Disease-Mediated Gingival Overgrowth

doi:

Figure Lengend Snippet: The effects of TG on the activity of cathepsins. Gingival fibroblasts were cultured with or without 100 ng/ml of nifedipine in the presence or absence of the indicated concentrations of Ca mobilizer TG. Twenty-four hours later, cellular proteins were extracted and were subjected to enzyme assay. TG suppressed both cathepsin-(B+L) and -B activity, while nifedipine alone specifically suppressed cathepsin-(B+L) activity. Values are expressed as the % activity against that of controls (day 0).

Article Snippet: Reagents The blocker of L-type Ca (2+) channels, nifedipine, was from Bayer AG Pharma Research Center (Wuppertal, Germany).

Techniques: Activity Assay, Cell Culture, Enzymatic Assay