Journal: Cells

Article Title: The Role of Two-Pore Channels in Norepinephrine-Induced [Ca 2+ ] i Rise in Rat Aortic Smooth Muscle Cells and Aorta Contraction

doi: 10.3390/cells8101144

Figure Lengend Snippet: Levels of TPC1 and TPC2 mRNA relative to β-actin mRNA ( A ). Western-blot of TPC1 and TPC2 proteins ( B ) isolated from rat aorta SMCs and relative levels of TPC1 and TPC2 ( C ). The values panel A are the means of four measurements in different SMCs preparation + SEM (* p < 0.01). In panel B, the typical image of Western blots from one of three different cell lysates is presented. (+) Marks hybridization in the presence of blocking peptide. In panel C, n = 3 and n.s.—not significant.

Article Snippet: cis -NED 19, trans -NED 19, U73122, and U73343 were acquired from Tocris (Bristol, UK); Fura2/AM, CalciumGreen/AM, and LysoTracker Red DND-99 from ThermoFischer Scientific (Waltham, MA, USA); Mitotracker Deep Red (M22426, Invitrogen), ER-tracker Red (E34250, Invitrogen), WGA Alexa Fluor 594 (Thermo Fisher, W11262), and antibodies anti-TPC1 (#ACC-071) and anti-TPC2 (#ACC-072) from Alomone Labs (Jerusalem, Israel); HRP-conjugated goat anti-rabbit IgG antibodies from BioRad (#170-6515); and the agonists of the receptors from SigmaAldrich (St. Louis, MO, USA).

Techniques: Western Blot, Isolation, Hybridization, Blocking Assay

Journal: Cells

Article Title: The Role of Two-Pore Channels in Norepinephrine-Induced [Ca 2+ ] i Rise in Rat Aortic Smooth Muscle Cells and Aorta Contraction

doi: 10.3390/cells8101144

Figure Lengend Snippet: Decrease in [Ca 2+ ] i rise in response to NE in SMCs transfected with siRNA against TPC1. ( A ) Kinetics of [Ca 2+ ] i rise in SMCs transfected with siRNA against TPC1 and nontarget siRNA. The curves from one of four transfection experiments are presented. Each point at the curves is an average of six parallel measurements. ( B ) Calcium responses to NE (100 μM) and angiotensin II (0.1 μM) in SMCs transfected with nontarget siRNA, and siRNA against TPC1 and against TPC2. [Ca 2+ ] i rise in SMCs transfected with nontarget siRNA is taken as 100%. The mean values + SEM from four independent transfection experiments with different SMCs preparations are presented. There is significant difference between the responses of SMCs transfected with anti-TPC1 siRNA and with nontarget (** p < 0.01) or anti-TPC2 siRNA (* p < 0.05).

Article Snippet: cis -NED 19, trans -NED 19, U73122, and U73343 were acquired from Tocris (Bristol, UK); Fura2/AM, CalciumGreen/AM, and LysoTracker Red DND-99 from ThermoFischer Scientific (Waltham, MA, USA); Mitotracker Deep Red (M22426, Invitrogen), ER-tracker Red (E34250, Invitrogen), WGA Alexa Fluor 594 (Thermo Fisher, W11262), and antibodies anti-TPC1 (#ACC-071) and anti-TPC2 (#ACC-072) from Alomone Labs (Jerusalem, Israel); HRP-conjugated goat anti-rabbit IgG antibodies from BioRad (#170-6515); and the agonists of the receptors from SigmaAldrich (St. Louis, MO, USA).

Techniques: Transfection