For supplemental staining, mouse anti-vinculin monoclonal antibody (Sigma) was used at 1∶40 dilution, after which cells were incubated with Alexa Fluor 488 goat anti-mouse (Invitrogen) at 1∶200 dilution.
Body weight and 24-hour food intake in uPAR-null mice were similar to wild-type mice (Supplemental Figure 3), and administration of CCK8s (2.5 nmol/kg ip) also inhibited food intake in uPAR−/− mice; however, PAI-1 (2.5 nmol/kg ip) alone had no effect on food intake and did not suppress the effect of CCK8s.
Interestingly, the ketone body content increased at 4 C to the same level as after overnight fasting period in control mice, whereas hadh−/− mice showed significantly lower levels than control mice (Supplemental Fig. 4).
We further corroborated these findings in the mouse (Supplemental Fig. 1).
Purity and viability of LSECs were up to 93.6 ± 1.7% and 88.4 ± 0.5% confirmed by CD146 + F4/80- and 7-AAD + flow cytometry analysis separately, and yield of LSECs was approximately (2.1 ± 0.2) × 106 per mouse, (Supplemental Fig. 1A C).
Specific PCR primers were designed according to the target gene sequences of mouse (Supplemental Table. 1).
As in the striatum, REST expression was increased in the whole brain of R6/2 mice, when compared to control mice (Supplemental figure S6).
Body weight, food intake, adiposity, liver weight were equivalent between Ob-CT mice and Ob-GLP-2 mice (supplemental data 3).
Importantly, however, in PAI-1−/− mice, the dose-response curve for inhibition of food intake by CCK8s was closely similar to that of C57BL/6 mice, exogenous PAI-1 increased food intake as in wild-type mice (Supplemental Figure 6), and the satiety response to CCK persisted in infected mice.
Lipid and protein metabolism, fatty acid beta-oxidation, cell death, apoptosis, peroxisome organization, and biogenesis were significantly upregulated in untreated db/db mice when compared to control db/+ mice (Supplemental Tables S1, S2, S3, S4).
Analysis of 4-hour fasted plasma samples at 24 weeks revealed no significant differences in sodium or potassium concentrations in male and female Crh− 120 /+ mice (Supplemental Table 1), but female Crh− 120 /+ mice may have been dehydrated, as indicated by the increased creatinine, total protein, albumin, and uric acid concentrations (Supplemental Table 1).
