Fig 1: 4-TBB is anticonvulsant in the mouse pentylenetetrazole (PTZ)-induced seizure model. (A,B) Exposure to 4-TBB increases time to onset of Straub tail (P=0.004) (A) and generalised tonic-clonic seizure (P=0.001) (B). (C). Post-mortem analysis of brains, taken from the mice used in the assay, shows that expression of PUM1 (P<0.0001) and PUM2 (P=0.01) is upregulated in animals treated with 4-TBB compared to controls (set as 1). Bars show means±s.d. Inset shows an example western blot. Significance was tested in A-C using unpaired two-tailed Student's t-tests. (D) Western blot analysis of Nav1.1, Nav1.2, Nav1.6 and GLUR2 expression in mouse brain shows that exposure to 4-TBB is sufficient to downregulate Nav1.1 (P=0.005), Nav1.2 (P=0.007) and GLUR2 (P=0.001), but not Nav1.6 (P=0.75). n=14. Bars show means±s.d. Significance was tested using one-way ANOVA [F(4,65)=6.7, P=0.0001] with correction for multiple comparisons (Dunnett's). Inset shows an example western blot. **P<0.01, ***P<0.001.
Fig 2: Characterisation of RAB216 shows it to be more potent than 4-TBB. (A) Western blot analysis of the effect of exposure of mouse brain to RAB216 on PUM1 (n=4, P=0.07) and PUM2 (n=4, P=0.01) expression (unpaired two-tailed Student's t-tests). (B) VPA, which is an effective anticonvulsant in this assay, had no effect on expression of PUM1 or PUM2 (n=4, P>0.05). (C,D) Effect of RAB216 on Straub tail (P=0.03) (C) and first appearance of a tonic-clonic seizure (P=0.01) in the PTZ-induced seizure assay (D). Timings were capped at 20 min (1200 s). n=15 (unpaired two-tailed Student's t-tests). (E) Percentage of animals exposed to PTZ showing tonic-clonic seizure following exposure to either saline (CTRL) or RAB216 (P=0.05) (n=15 and n=14, respectively, Fisher’s exact test). (F) Exposure to RAB216 reduces 6 Hz electrically induced seizure duration (n=9, P=0.013). Levetiracetam (LEV; 100 mg/kg, n=9, P<0.0001) was used as a positive control with known efficacy in this assay. Significance was tested using one-way ANOVA [F(2,24)=16.88] with correction for multiple comparisons (Dunnett's). *P<0.05, **P<0.01, ***P<0.001.
Supplier Page from Abcam for Anti-Pumilio 2 antibody