Figure 1. IKKε and TBK1 overexpression decrease sensitivity to the β-adrenergic/cAMP pathway in 3T3-L1 adipocytes.

(A) Fold increase in Ucp1 expression in 3T3-L1 adipocytes expressing empty vector, Flag-IKKε, or Flag-IKKε K38A following treatment with or without 10 μM ISO (black bars) or 10 μM CL-316,243 (CL, gray bars) for 4 hr. **p<0.01. Performed in triplicate. (B) Glycerol release from 3T3-L1 adipocytes expressing empty vector (white bars), Flag-IKKε (black bars), or Flag-IKKε K38A (gray bars) treated with or without 10 μM ISO or 10 μM CL. *p<0.05 and **p<0.01. Performed in triplicate. (C) Immunoblots of whole cell lysates from Figure 1B. Results were replicated in triplicate. D.E. stands for dark exposure and L.E. stands for light exposure. (D) Immunoblots of whole cell lysates from 3T3-L1 adipocytes expressing empty vector or Flag-IKKε treated with or without 50 μM FSK for 15 min. Results were replicated in multiple experiments. (E) cAMP levels from 3T3-L1 adipocytes expressing empty vector, Flag-IKKε, or Flag-IKKε K38A treated with or without 10 μM ISO or 50 μM FSK for 15 min. **p<0.0001 and *p<0.05. Performed in triplicate.

DOI: http://dx.doi.org/10.7554/eLife.01119.003

Figure 1—figure supplement 1. IKKε and TBK1 overexpression decrease sensitivity to the β-adrenergic/cAMP pathway in 3T3-L1 adipocytes.

(A) Immunoblots of whole cell lysates from 3T3-L1 adipocytes expressing empty vector, Flag-IKKε, or Flag-IKKε K38A treated with or without 10 μM ISO for 15 min. Results were replicated in multiple experiments. (B) Immunoblots of whole cell lysates from 3T3-L1 adipocytes expressing increasing amounts of Flag-IKKε or Flag-TBK1 treated with or without 10 μM ISO (top panel) or 50 μM FSK (bottom panel) for 15 min. Results were replicated in multiple experiments.

DOI: http://dx.doi.org/10.7554/eLife.01119.004