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. 2021 Oct 11;9:737872. doi: 10.3389/fcell.2021.737872

FIGURE 3.

FIGURE 3

Irisin dependent CXCL1 release was stimulated from differentiating subcutaneous (SC) and deep-neck (DN) area adipocytes. SC and DN preadipocytes were differentiated and treated as in Figures 1, 2. Where indicated, irisin was omitted from the differentiation medium at day 14. Conditioned differentiation media was collected and secreted CXCL1 was measured by sandwich ELISA. (A) Quantification of CXCL1 gene expression as determined by RNA Sequencing (n = 9) or RT-qPCR (B) normalized to GAPDH (n = 5), (C) CXCL1 release by ex vivo differentiating SC and DN adipocytes into the conditioned media collected at the indicated intervals, in the presence or absence of irisin (n = 4), (D) CXCL1 release in conditioned medium collected at indicated intervals from untreated (21 days) and irisin treated (14 and 21 days as indicated) cell-culture samples (n = 3), (E) CXCL1 release from differentiating adipocytes with or without irisin treatment, in the presence or absence of 10 μg/ml RGDS (n = 4). Comparisons are for the respective days in case of ELISA. Data presented as Mean ± SD. ∗p < 0.05, ∗∗p < 0.01. Statistics: GLM (A), One-way ANOVA with Tukey’s post-test (B), Two-way ANOVA with Tukey’s post-test (C–E).