Increased expression level of ANGPTL8 in white adipose tissue under acute and chronic cold treatment

Fig. 1. Animal weight, food consumed, and adipose tissue (SAT and BAT) weight at different time points of cold exposure (Day0 (Control-no cold exposure), Day1, Day3, Day 5, and Day10). A: Animal weight (g) at different days of cold treatment. B: Amount of food consumed daily (g) over the ten days of cold treatment. C: SAT weight (mg) at different days of cold treatment. D: BAT weight (mg) different days of cold treatment. * indicates P ≤ 0.05; *** indicates P ≤ 0.0001 as determined using Student’s t-test

Fig. 2. Liver ANGPTL3, ANGPTL4 and ANGPTL8 gene expression measured at Day 0 (Control no cold exposure, n = 5), Days 1, 3, 5 and 10 post cold treatment at 4 0 C (n = 10 each). A: Expression level of ANGPTL3 in Liver as measured by RT-PCR at different time points of cold treatment. B: Expression level of ANGPTL4 in liver as measured by RT-PCR at different time points of cold treatment. C: Gene expression level of ANGPTL8 as measured by RT-PCR at different time points of cold treatment. ns indicates no significance; * indicates P ≤ 0.05; ** indicates P ≤ 0.001, as determined using Student’s t-test

Fig. 3. SAT UCP1, ANGPTL4 and ANGPTL8 gene expression measured at Day 0 (Control no cold exposure, n = 5), Days 1, 3, 5 and 10 post cold treatment at 4 0 C (n = 10 each). A: Gene expression level of ANGPTL4 in SAT as measured by RT-PCR at different time points of cold treatment. B: Gene expression level of ANGPTL8 in SAT as measured by RT-PCR at different time points of cold treatment. C: Gene expression level of UCP1 in SAT as measured by RT-PCR at different time points of cold treatment. D: Gene expression level of LPL in SAT as measured by RT-PCR at different time points of cold treatment. E: Gene expression level of ATGL in SAT as measured by RT-PCR at different time points of cold treatment. * indicates P-value ≤ 0.05; ** indicates P ≤ 0.001; *** indicates P ≤ 0.0001 as determined using Student’s t-test

Fig. 4. BAT UCP1, ANGPTL4 and ANGPTL8 gene expression measured at Day 0 (Control no cold exposure, n = 5), Days 1, 3, 5 and 10 post cold treatment at 4 0 C (n = 10 each). A: Gene expression level of ANGPTL4 in BAT as measured by RT-PCR at different time points of cold treatment. B: Gene expression level of ANGPTL8 in BAT as measured by RT-PCR a at different time points of cold treatment. C: Gene expression level of UCP1 in BAT as measured by RT-PCR at different time points of cold treatment. D: Gene expression level of LPL in BAT as measured by RT-PCR at different time points of cold treatment. E: Gene expression level of ATGL in BAT as measured by RT-PCR at different time points of cold treatment. * indicates P-value ≤ 0.05; ns indicates no significance; ** indicates P ≤ 0.001; *** indicates P ≤ 0.0001 as determined using Student’s t-test

Fig. 5. Western blotting analysis of ANGPTL8 protein level in SAT, BAT and Liver measured at Day 0 (Control no cold exposure), Days 1, 5 and 10 post cold treatment at 4 0 C (n = 2 for all groups). A: ANGPTL8 protein expression in SAT at different time points of cold treatment. B: ANGPTL8 protein expression in BAT at different time points of cold treatment. C: ANGPTL8 protein expression in Liver at different time points of cold treatment

Fig. 6. Representative Immunohistochemistry staining of ANGPTL8 protein level in SAT, BAT and Liver measured at Day 0 (Control no cold exposure), Days 1, 3, 5 and 10 post cold treatment at 4 0 C (n = 3 for all groups). A: ANGPTL8 protein expression in SAT at different time points of cold treatment. B: ANGPTL8 protein expression in BAT at different time points of cold treatment. C: ANGPTL8 protein expression in Liver at different time points of cold treatment