Figure 5: IL-1β-induced Vegfa expression in adipocytes.

(a) Vegfa expression is increased in non-myeloid cells under obesity. CD11b⁺ and CD11b⁻ cells were purified by magnetic beads in tumours from ND or DIO mice. Data represent the mean fold change compared with ND±s.d. Expression is relative to Actb (n=3 for each group). (b,c) Indicated cells were left untreated (cont), treated with 100 ng ml⁻¹ rIL-1β or CM from non-treated or 100 ng ml⁻¹ rIL-1β-treated primary BMDM. Data represent the average mRNA expression of Vegfa relative to Ppia as fold change compared with cont±s.d. (d) Adipocytes were treated as indicated with 100 ng ml⁻¹ rIL-1β, 5 μM NFκB inhibitor (BMS345541) and 40 μM JNK inhibitor. Data represent the average mRNA expression of Vegfa relative to Ppia±s.d. (n=3 for all groups). (e) Primary mammary adipocytes were treated with 100 ng ml⁻¹ rIL-1β for the indicated time, and the indicated proteins were separated by SDS–PAGE followed by immunoblotting with the indicated antibodies. All studies represent results from two to three repeats. For all panels, the group means were compared by Student’s t-test to determine significance.