Fig 1: Young NHP brain has no sex bias in PON2 isoforms expression in STR and SN regions (n = 5). Protein expression of PON2 isoforms, i.e., 39 kDa and 41 kDa in STR (A, B) and SN (C, D), respectively. Representative blot showing PON2 isoforms expression in STR (E), and SN (F). Corresponding image of total protein in STR (G) and SN (H). Optical density of UCP5 and UCP4 bands were normalized to total protein per lane. The normality of infant male and female data was confirmed by a Shapiro–Wilk test. 39 kDa PON2 STR Male (W = 0.97, p = 0.89) Female (W = 0.91, p = 0.50); 41 kDa PON2 Male (W = 0.96, p = 0.87) Female (W = 0.80, p = 0.09); 39 kDa PON2 Male (W = 0.84, p = 0.18) Female (W = 0.83, p = 0.15); 41 kDa PON2 SN Male (W = 0.97, p = 0.87) Female (W = 0.97, p = 0.90). Data were expressed as mean ± SEM. Male and female data in each region was compared by two-tailed unpaired Student's t test. 39 kDa PON2 STR (two-tailed t (8) = 0.07, p = 0.94), 41 kDa PON2 STR (two-tailed t (8) = 0.0, p > 0.99), 39 kDa PON2 SN (two-tailed t (8) = 0.03, p = 0.97), 41 kDa PON2 SN (two-tailed t (8) = 0.0, p > 0.99)
Fig 2: Impact of 1-week and 3-week PIO treatment on UCP-4 and UCP-5 protein expression in STR and SN of adult male NHPs (n = 5). Protein expression of UCP4 and UCP5 in STR (A, B) and SN (C, D), respectively. Representative blot showing UCP4 and UCP5 expression in STR (E, G), and SN (F, H) region. Corresponding image of total protein in STR (I) and SN (J). Optical density of UCP5 and UCP4 bands were normalized to total protein per lane. Optical density of UCP5 and UCP4 bands were normalized to total protein per lane. The normality of data in each group was confirmed by a Shapiro–Wilk test. UCP4 STR VEH (W = 0.88, p = 0.32), PIO 1 week (W = 0.95, p = 0.78), PIO 3 weeks (W = 0.84, p = 0.16); UCP5 STR VEH (W = 0.91, p = 0.51), PIO 1 week (W = 0.96, p = 0.81), PIO 3 weeks (W = 0.81, p = 0.10); UCP4 SN VEH (W = 0.87, p = 0.27), PIO 1 week (W = 0.87, p = 0.27), PIO 3 weeks (W = 0.93, p = 0.61); UCP5 SN VEH (W = 0.91, p = 0.51), PIO 1 week (W = 0.88, p = 0.34), PIO 3 weeks (W = 0.80, p = 0.08). Data were expressed as mean ± SEM. Data from different groups in each region were first analyzed for homogeneity of variance using Brown–Forsythe test (UCP4 STR: F (2, 12) = 1.551, p = 0.2518), UCP5 STR: F (2, 12) = 1.492, p = 0.2649); UCP4 SN: F (2, 12) = 1.593, p = 0.2435), UCP5 SN: F (2, 12) = 2.097, p = 0.1655)) and then compared by one-way ANOVA followed by Tukey HSD post-hoc test for multiple comparisons. UCP4 STR; one-way ANOVA: F (2, 12) = 9.91, p = 0.0029), UCP5 STR; one-way ANOVA: F (2, 12) = 12.94, p = 0.0010); UCP4 SN; one-way ANOVA: F (2, 12) = 22.47, p < 0.0001), UCP5 SN; one-way ANOVA: F (2, 12) = 5.06, p < 0.025). Asterisks indicate statistical significance in comparison with vehicle group, *p < 0.05, **p < 0.01
Fig 3: Adult NHP brain has sex bias in PON2 isoforms expression in STR and SN regions. STR (n = 5) and SN (n = 4). Protein expression of PON2 isoforms, i.e., 39 kDa and 41 kDa in STR (A, B) and SN (C, D), respectively. Representative blot showing PON2 isoforms expression in STR (E), and SN (F). Corresponding image of total protein in STR (G) and SN (H). Optical density of UCP5 and UCP4 bands were normalized to total protein per lane. The normality of infant male and female data was confirmed by a Shapiro–Wilk test. 39 kDa PON2 STR Male (W = 0.84, p = 0.17) Female (W = 0.97, p = 0.87); 41 kDa PON2 Male (W = 0.95, p = 0.77) Female (W = 0.92, p = 0.54); 39 kDa PON2 Male (W = 0.92, p = 0.58) Female (W = 0.91, p = 0.51); 41 kDa PON2 SN Male (W = 0.87, p = 0.33) Female (W = 0.95, p = 0.73). Data were expressed as mean ± SEM. Male and female data in each region was compared by two-tailed unpaired Student's t test. 39 kDa PON2 STR (two-tailed t (6) = 3.75, p = 0.0095), 41 kDa PON2 STR (two-tailed t (8) = 4.87, p = 0.0012), 39 kDa PON2 SN (two-tailed t (8) = 0.03, p = 0.97), 41 kDa PON2 SN (two-tailed t (6) = 2.72, p = 0.034). Asterisks indicate statistical significance in comparison with vehicle group, *p < 0.05, **p < 0.01, ***p < 0.001
Fig 4: Adult NHP brain has sex bias in UCP-4 and UCP-5 expression in SN but not in STR region. STR (n = 5) and SN (n = 4). Protein expression of UCP4 and UCP5 in STR (A, B) and SN (C, D), respectively. Representative blot showing UCP4 and UCP5 expression in STR (E, G), and SN (F, H) region. Corresponding image of total protein in STR [I] and SN [J]. Optical density of UCP5 and UCP4 bands were normalized to total protein per lane. Optical density of UCP5 and UCP4 bands were normalized to total protein per lane. The normality of infant male and female data was confirmed by a Shapiro–Wilk test. UCP4 STR Male (W = 0.80, p = 0.085) Female (W = 0.91, p = 0.52); UCP5 STR Male (W = 0.82, p = 0.13) Female (W = 0.95, p = 0.79); UCP4 SN Male (W = 0.85, p = 0.23) Female (W = 0.90, p = 0.46); UCP5 SN Male (W = 0.99, p = 0.99) Female (W = 0.99, p = 0.97). Data were expressed as mean ± SEM. Male and female data in each region was compared by two-tailed unpaired Student's t test. UCP4 STR (two-tailed t (8) = 0.84, p = 0.93); UCP5 STR (two-tailed t (8) = 1.78, p = 0.11); UCP4 SN (two-tailed t (6) = 5.57, p = 0.0014); UCP5 SN (two-tailed t (6) = 5.87, p = 0.0011). Asterisks indicate statistical significance in comparison with vehicle group, *p < 0.05, **p < 0.01
Fig 5: Young NHP brain has no sex bias in UCP4 and UCP5 expression in STR and SN regions (n = 5). Protein expression of UCP4 and UCP5 in STR (A, B) and SN (C, D), respectively. Representative blot showing UCP4 and UCP5 expression in STR (E, G), and SN (F, H) region. Corresponding image of total protein in STR (I) and SN (J). Optical density of UCP5 and UCP4 bands were normalized to total protein per lane. The normality of Infant male and female data was confirmed by a Shapiro–Wilk test. UCP4 STR Male (W = 0.86, p = 0.25) Female (W = 0.96, p = 0.81); UCP5 STR Male (W = 0.78, p = 0.057) Female (W = 0.84, p = 0.18); UCP4 SN Male (W = 0.93, p = 0.65) Female (W = 0.86, p = 0.25); UCP5 SN Male (W = 0.88, p = 0.35) Female (W = 0.93, p = 0.64). Data were expressed as mean ± SEM. Male and female data in each region was compared by two-tailed unpaired Student's t test. UCP4 STR (two-tailed t (8) = 0.99, p = 0.34), UCP5 STR (two-tailed t (8) = 1.85, p = 0.10), UCP4 SN (two-tailed t (8) = 0.73, p = 0.48), UCP5 SN (two-tailed t (8) = 2.23, p = 0.055)
Supplier Page from Abcam for Anti-UCP4 antibody