The liver X receptors (LXR)α and LXRβ are transcription factors belonging to the nuclear receptor family which play a central role in metabolic homeostasis being master regulators of key target genes in the glucose and lipid pathways. Wild-type (Wt), LXRα^-/- and LXRβ^-/- mice were fed a chow diet with (treated) or without (control) the synthetic dual LXR agonist GW3965 for 5 weeks. GW3965 raised intra-hepatic triglyceride (Tg) level but, surprisingly, reduced serum Tg level through the activation of serum lipase activity. The serum Tg reduction was associated with a repression of both catecholamine-stimulated lipolysis and relative glucose incorporation into lipid in isolated adipocytes through activation of LXRβ. We also demonstrated that LXRα is required for basal (non-stimulated) adipocyte metabolism while LXRβ acts as a repressor of lipolysis. On the contrary, in skeletal muscle (SM) the lipogenic and the cholesterol transporters LXR target genes were markedly induced in Wt and LXRα^-/- mice and to a lower extent in LXRβ^-/- mice, following treatment with GW3965. Moreover, Tg content was reduced in SM of LXRβ^-/- mice associated with an increased expression of the main Tg-lipase genes Hsl and Atgl. Energy expenditure was increased and a switch from glucose to lipid oxidation was observed. In conclusion, we provide evidence that LXR might be an essential regulator of the lipid balance between tissues to ensure appropriate control of the flux of fuel. Importantly, we show that after chronic treatment with GW3965, SM becomes the target tissue for LXR activation, as opposed to liver in acute treatment.