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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