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).

Techniques:

Journal: PLoS ONE

Article Title: Serum response factor regulates smooth muscle contractility via myotonic dystrophy protein kinases and L-type calcium channels

doi: 10.1371/journal.pone.0171262

Figure Lengend Snippet: (A) Expression levels of L-type calcium channel subtypes in SMC of jejunum and colon. (B) Expression levels of Cacna1c variants in SMC and tissue of jejunum and colon. (C) A genomic map of Cacna1c variants. Five variable regions (V1-5) are indicated. Exons are numbered 1–48. (D) Magnified view of variable regions showing alternatively started or spliced exons (indicated as exon numbers). Seven exons containing alternative transcriptional start sequence are shown by a star (*). Long (L) and short (S) exons that are differentially started or spliced are indicated.

Article Snippet: Primary antibodies against the following antigens were used: CACNA1C (guinea pig, 1:1000, Alomone Labs, Jerusalem, Israel), SRF (rabbit, 1:500, Santa Cruz Biotechnology, Dallas, TX), or GAPDH (rabbit, 1:1500, Cell Signaling Technology, Danvers, MA).

Techniques: Expressing, Sequencing

Journal: PLoS ONE

Article Title: Serum response factor regulates smooth muscle contractility via myotonic dystrophy protein kinases and L-type calcium channels

doi: 10.1371/journal.pone.0171262

Figure Lengend Snippet: (A) PCR validation of alternatively started and spliced exons of Cacna1c in WT and KO jejunum smooth muscle at KO days 5, 10, and 15. NTC is non template control. Primer sets were designed from variant exons in the regions (e.g. E1-3, forward primer spanning a region on exon 1 and reverse primer on exon 3). (B) qPCR data showing decreased expression of Cacna1c variants starting at exon 1 and exon 2 long and short forms (E1-3, E2L-3, and E2L/S-3) at KO days 5, 10, 15, and 20. E1-3, a region spanning exons 1 and 3; E2L-3, a region spanning exon 2 long (L) form and exon 3; E2L/S-3, a region spanning exon 2 long (L) and short (S) forms and exon 3. (C) Western blot showing decreased levels of CACNA1C protein in Srf KO muscle. (D) Consensus sequence of 5’ splice donor and 3’ splice acceptor sites. (E) A topological map of CACNA1C variants. Amino acid sequence is written in small circles. Four motifs are indicated as I-IV and six transmembrane domains, S1-S6. Four pore regions are also indicated. Colors on amino acid sequence show particular regions and domains: red, missing or inserted peptides from differentially spliced exons; purple, voltage sensors in S4 transmembrane domains; green, start codons found in differentially spliced variants (*, start codons deduced from indicated exons that are differentially spliced; blue, β subunit binding domain; brown, CaM (calmodulin) binding domain; orange, PKA (protein kinase A) phosphorylation site. Alignment of alternatively spliced exons E9/10, E24/25, and E32/33 are shown. (F) Isometric force recordings from antrum and colon of WT and KO mice. Bay K8644 (1 μM) and high potassium (K + ) Krebs (36 mM and 72 mM) were applied to the tissues (indicated by bar and arrows). (G) The graph summarizes the results for 9 antral and 5 colonic WT and KO tissues. The responses to Bay K8644, 36 mM K + , and 72 mM K + were significantly decreased in KO antrums, and the responses to 36 mM K + and 72 mM K + were significantly reduced in KO colons compared to WT. * and ** represent p ≤ 0.05 and p ≤ 0.01 respectively.

Article Snippet: Primary antibodies against the following antigens were used: CACNA1C (guinea pig, 1:1000, Alomone Labs, Jerusalem, Israel), SRF (rabbit, 1:500, Santa Cruz Biotechnology, Dallas, TX), or GAPDH (rabbit, 1:1500, Cell Signaling Technology, Danvers, MA).

Techniques: Variant Assay, Expressing, Western Blot, Sequencing, Binding Assay

Journal: PLoS ONE

Article Title: Serum response factor regulates smooth muscle contractility via myotonic dystrophy protein kinases and L-type calcium channels

doi: 10.1371/journal.pone.0171262

Figure Lengend Snippet: Model showing the possible molecular pathways by which SRF regulates contractility via DMPK and CACNA1C in SMC.

Article Snippet: Primary antibodies against the following antigens were used: CACNA1C (guinea pig, 1:1000, Alomone Labs, Jerusalem, Israel), SRF (rabbit, 1:500, Santa Cruz Biotechnology, Dallas, TX), or GAPDH (rabbit, 1:1500, Cell Signaling Technology, Danvers, MA).

Techniques: