Figure 5.

Obese mice treated with liposomal-CGS or adenosine injections exhibited elevated levels of total FoxO1 and FoxO3 with increased nuclear localization in both and reduced levels of inactive phospho-FoxO1 compared to obese controls. (A) 40 × hpf magnified joint sections stained for active FoxO1 (green fluorescence) using IF/IHC in the obesity mouse model of arthritis representative from 3 mice per group. A 1-way ANOVA (p < 0.0001) was performed assessing individual cell thresholded fluorescence to calculate the percent of IHC 40 × HPF in 3 mice per group by counting medial compartment intra-articular chondrocyte cell fluorescence. Mean percent and standard deviation for the 5 in vivo groups are recorded in order by non-obese CTRL, obese CTRL, liposome only joint injection, adenosine injection, and A2AR agonist injection (52% ± 2.9%, 29% ± 4.3%, 32% ± 6.2%, 72% ± 4.5%, 70% ± 5.0%; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, n = 3, 1-way ANOVA followed by individual comparisons using Tukey multiple comparison tests). (B) 40 × hpf magnified joint sections stained for active FoxO3 (green fluorescence) using IF/IHC in the obesity mouse model of arthritis representative from 3 mice per group. A 1-way ANOVA (p < 0.0001) was performed assessing individual cells thresholded by level/location of fluorescence was employed to calculate the percent of fluorescence per cell in IHC sections as measured from a representative 40 × HPF for sections obtained from each mice in each treatment group. Mean percent and standard deviation for the 5 in vivo groups are recorded in order by non-obese CTRL, obese CTRL, liposome only joint injection, adenosine injection, and A2AR agonist injection (52% ± 5.4% ± 24% ± 7.8%, 34% ± 3.0%, 70% ± 3.3%, 70% ± 2.3%; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, n = 3, 1-way ANOVA followed by individual comparisons using Tukey multiple comparison tests). Figure labels: F is femur, T is tibia, M is meniscus.