Journal: eLife

Article Title: Mutations in L-type amino acid transporter-2 support SLC7A8 as a novel gene involved in age-related hearing loss

doi: 10.7554/eLife.31511

Figure Lengend Snippet: Hearing phenotype of C57BL6/J-129Sv Slc7a8 knockout mice. ( A ) Representative image of western bloting of total membranes from kidney, brain and intestine of wild-type (+/+) and Slc7a8 knock out (-/-) mice in the absence (-) or presence (+) of 100 mM dithiothreitol reducing agent (DTT) of three independent biological samples for both sexes (male and female). Protein (50 μg) were loaded in 7% acrylamide SDS-PAGE gel. Molecular mass standard (KDa) are indicated. Red arrows point SLC7A8/CD98hc heterodimer band as well as the light subunit SLC7A8. Upper panel: Rabbit anti-SLC7A8. Bottom panel: Mouse anti-βactin. ( B ) Pre-Pulse Inhibition of the acoustic startle response (PPI). Mean and SEM are represented. Pulse: 120 dB single pulse. Pre-pulse inhibition test: six different pre-pulse intensities (70 to 90 dB) in pseudo random order with 15 s inter-trial intervals. Wild type (white circles, n = 19) and Slc7a8 −/− (blue circles, n = 15) from 4- to 7-month-old are represented. Significant differences were determined using paired Student’s t-test, ***p

Article Snippet: For specific SLC7A8 light subunit detection, samples were run in the presence of 100 mM of dithiothreitol (SigmaAldrich Ref:D9779).

Techniques: Knock-Out, Mouse Assay, Western Blot, SDS Page, Inhibition

Journal: Molecular Pain

Article Title: Polysulfide evokes acute pain through the activation of nociceptive TRPA1 in mouse sensory neurons

doi: 10.1186/s12990-015-0023-4

Figure Lengend Snippet: Involvement of the N-terminal cysteine residues of mouse TRPA1 in its activation by polysulfide. (A) The Na 2 S 3 (10 μM)-induced [Ca 2+ ] i increase was inhibited by dithiothreitol (DTT) (a) 2 min before and during 4 min application of Na 2 S 3 , (b) after 4 min in HEK 293 cells expressing mouse TRPA1 (mTRPA1-HEK). The upper panels show [Ca 2+ ] i responses to Na 2 S 3 without DTT, and the lower ones those in the presence of DTT. (B) Summarized effects of DTT. (a) Open and filled columns show the increases of [Ca 2+ ] i responses to Na 2 S 3 in the absence (Control) and presence of DTT, respectively. (b) Times required for half-decline of [Ca 2+ ] i responses to Na 2 S 3 (T 1/2 ) in the absence (Control) and presence of DTT. T 1/2 was calculated by subtracting the value of the time when the Na 2 S 3 -induced [Ca 2+ ] i increase was reduced by half from that when Na 2 S 3 -induced [Ca 2+ ] i increase peaked. Columns with vertical lines show mean ± SEM (a; n = 23–32, b; n = 55–63, from three different transfections). **P

Article Snippet: Dithiothreitol (DTT, DW, 1 M), polysulfides (Na2 S3 and Na2 S4 ), and ruthenium red (DW, 0.01 M) were from Wako, Osaka, Japan.

Techniques: Activation Assay, Expressing, Transfection