Article Title: Neuromedin B receptor stimulation of Cav3.2 T-type Ca2+ channels in primary sensory neurons mediates peripheral pain hypersensitivity
Figure Lengend Snippet: Activation of NmbR stimulates recombinant Cav3.2 channels heterologously expressed in HEK293 cells. A, western blot analysis of NmbR in HEK293 cells transiently transfected with NmbR cDNA. Blots depicted are representative of three independent experiments. B, membrane localization of NmbR in transfected HEK293 cells. Alphabets a through c in the diagram indicate the differential interference contrast (DIC, a ), the EGFP fluorescent signals of NmbR ( b ), and the merged image ( c ), respectively. C, exemplary current traces show the effect of Nmb (100 nM) on Cav3.1 (α1G), Cav3.2 (α1H) and Cav3.3 (α1I) channel currents. Currents were elicited by a 100 ms depolarizing step pulse from the holding potential of -110 mV to -30 mV. D, summary data show the effect of 100 nM Nmb on Cav3.1 ( n = 10), Cav3.2 ( n = 9) and Cav3.3 ( n = 7) channel currents. ** p
Article Snippet: The membranes were blocked with 5% skim milk in Tris-buffered saline containing 0.1% Tween-20 (TBST) and probed with antibodies against Nmb (rabbit, 1:1000, Abcam, Cat. No. ab191499), Cav3.1 (rabbit, 1:1000, Alomone, Cat. No. ACC-021), Cav3.2 (mouse, 1:1000, Novus, Cat. No. NBP1-22444), Cav3.3 (rabbit, 1:1000, Alomone, Cat. No. ACC-009), NmbR (rabbit, 1:500, Sigma, Cat. No. SAB4502914), phospho-AMPKα1 (rabbit, 1:500, Abcam, Cat. No. ab92701), AMPKα1 (rabbit, 1:600, Abcam, Cat. No. ab110036), Gαq/11 (mouse, 1:600, Santa Cruz Biotechnology, Cat. No. SC-365906), and Gβ (mouse, 1:500, Santa Cruz Biotechnology, Cat. No. SC-515822).
Techniques: Activation Assay, Recombinant, Western Blot, Transfection