Supplemental Figure 6

Supplemental Figure 6. Time spent freezing during a five-minute exposure to TMT. A history of CFA injury increased time spent freezing, and this increase was prevented by administration of HC30031. Data presented as mean value +/- SEM.

res <- aov(Perc ~ CFA * Drug, data = b)
summary(res)

##             Df Sum Sq Mean Sq F value   Pr(>F)    
## CFA          1    357   357.1    3.97 0.050885 .  
## Drug         1   1898  1898.5   21.10 2.29e-05 ***
## CFA:Drug     1   1091  1091.4   12.13 0.000932 ***
## Residuals   60   5398    90.0                     
## ---
## Signif. codes:  0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1

b %>%
  group_by(CFA) %>%
  pairwise_t_test(Perc ~ Drug)

## # A tibble: 2 × 10
##   CFA   .y.   group1 group2       n1    n2       p p.signif   p.adj p.adj.signif
## * <fct> <chr> <chr>  <chr>     <int> <int>   <dbl> <chr>      <dbl> <chr>       
## 1 Naive Perc  DMSO   HC-030031    16    16 4.65e-1 ns       4.65e-1 ns          
## 2 CFA   Perc  DMSO   HC-030031    16    16 1.04e-6 ****     1.04e-6 ****

b %>%
  group_by(Drug) %>%
  pairwise_t_test(Perc ~ CFA)

## # A tibble: 2 × 10
##   Drug     .y.   group1 group2    n1    n2       p p.signif   p.adj p.adj.signif
## * <fct>    <chr> <chr>  <chr>  <int> <int>   <dbl> <chr>      <dbl> <chr>       
## 1 DMSO     Perc  Naive  CFA       16    16 9.88e-4 ***      9.88e-4 ***         
## 2 HC-0300… Perc  Naive  CFA       16    16 2.69e-1 ns       2.69e-1 ns

TRPA-1 antagonism reduced freezing in CFA-primed mice, but had no effect on freezing during TMT for naive mice (F_1,60 = 12.13, p < 0.001). In the DMSO group, CFA-primed mice froze more than injury-naive mice (p < 0.001). TRPA-1 antagonism blocked the enhancing effect of a history of injury on freezing during TMT presentation. HC30031 treatment reduced freezing among CFA-primed mice (p < 0.001) such that freezing levels were not different from naive mice injected with HC30031 (p = 0.27). There was no effect of HC30031 administration on time spent freezing during TMT for injury-naive mice (p = 0.46).