Journal: Molecular Pain

Article Title: Role of anoctamin-1 and bestrophin-1 in spinal nerve ligation-induced neuropathic pain in rats

doi: 10.1186/s12990-015-0042-1

Figure Lengend Snippet: Bestrophin-1 is expressed in the spinal cord and DRG. RT-PCR ( a , b ) and western blot ( c , d ) analysis of bestrophin-1 (Best-1) at the ipsilateral dorsal portion of the spinal cord (SC) and DRG obtained from naïve (Na), sham (S) and spinal nerve ligated (SNL) rats. Insets show a representative band of the PCR product ( upper panels ) and immunoblot ( lower panels ) obtained with the specific bestrophin-1 and β-actin primers or antibodies, respectively. Data were normalized against β-actin and are expressed as the mean ± SEM of three independent rats. Note that spinal nerve injury did not modify Best-1 mRNA and protein expression.

Article Snippet: Membranes were blocked with 5% non-fat milk in phosphate-buffered saline at pH 7.4 containing (in mM) (137 NaCl, 2.7 KCl, 10 Na 2 HPO 4 and 2 KH 2 PO 4 ) with Tween 0.05% and they were incubated at 4°C overnight with rabbit anti-bestrophin-1 (ABC-001, 1:200; Alomone Labs, Jerusalem, Israel) or rabbit anti-anoctamin-1 (ACL-011, 1:100; Alomone Labs, Jerusalem, Israel).

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing

Journal: Molecular Pain

Article Title: Role of anoctamin-1 and bestrophin-1 in spinal nerve ligation-induced neuropathic pain in rats

doi: 10.1186/s12990-015-0042-1

Figure Lengend Snippet: Intrathecal administration of selective antibody anti-bestrophin-1 or anti-anoctamin-1 reverses established allodynia and reduces protein expression. Time-course of the antiallodynic effect of antibodies against bestrophin-1 (Best-1 ab, 2 μg/6 h × 30 h; a ) or anoctamin-1 (Ano-1 ab, 2 μg/6 h x 30 h; d ) in rats subjected to L5/L6 spinal nerve ligation. Paw withdrawal threshold was assessed every 6 h starting 12 days after spinal nerve ligation. Data are presented as the mean ± SEM for six animals. *Significantly different from the vehicle group (p < 0.05), as determined by two-way ANOVA followed by the Bonferroni test. Western blot analysis of bestrophin-1 (Best-1, b , c ) or anoctamin-1 (Ano-1, e , f ) at the ipsilateral dorsal portion of the spinal cord (SC) and DRG obtained from neuropathic rats with repeated intrathecal administration of antibodies against bestrophin-1 (Best-1 ab) or anoctamin-1 (Ano-1 ab). Data were normalized against β-actin and are expressed as the mean ± SEM of three independent rats. Insets in b – f show a representative blot obtained with bestrophin-1, anoctamin-1 and β-actin primary antibodies. *Significantly different from the vehicle (V) group (p < 0.05), as determined by the Student t test.

Article Snippet: Membranes were blocked with 5% non-fat milk in phosphate-buffered saline at pH 7.4 containing (in mM) (137 NaCl, 2.7 KCl, 10 Na 2 HPO 4 and 2 KH 2 PO 4 ) with Tween 0.05% and they were incubated at 4°C overnight with rabbit anti-bestrophin-1 (ABC-001, 1:200; Alomone Labs, Jerusalem, Israel) or rabbit anti-anoctamin-1 (ACL-011, 1:100; Alomone Labs, Jerusalem, Israel).

Techniques: Expressing, Ligation, Western Blot