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Fig. 7 | Journal of Neuroinflammation

Fig. 7

From: Polyunsaturated fatty acid receptors, GPR40 and GPR120, are expressed in the hypothalamus and control energy homeostasis and inflammation

Fig. 7

Lentiviral targeting of GPR120. In a, the mHypoA 2/29 CLU189 cell line was transfected with lentivirus containing five distinct (LV1–LV5) sequences of shRNA targeting GPR120 and, in addition, a scrambled control. Protein extracts were prepared and subjected to separation by SDS-PAGE, then transferred to a nitrocellulose membrane and blotted with anti-GPR120 antibody. The sequences LV1 and LV2 were utilized in in vivo experiments and were injected bilaterally into the arcuate nucleus (b). 7 days after LV1 or LV2 injection, hypothalami were dissected and used for protein extraction. Samples were subjected to separation by SDS-PAGE, transferred to nitrocellulose membranes and immunoblotted (IB) with an antibody against GPR120 (c). Swiss mice were fed for 4 weeks on a high-fat diet (HFD) and then injected with LV1 or the scrambled control construct into the arcuate nucleus bilaterally; after 10 weeks, the mice were subjected to intracerebroventricular cannulation and following a recovery period of 7 days, subjected to treatment with GW9508 (2.0 μl, 1.0 mM) twice a day for 6 days (d). Body mass (e and f) and caloric intake (g) were determined during the 10 weeks that followed LV1/scramble injection (before GW9508 treatment). Body mass (h), caloric intake (i), and energy efficiency (j) were evaluated during the 6 days that followed GW9508 treatment. At the end of the experimental period, IL10 protein expression was determined in extracts from the hypothalamus (k). In c and k, loading controls were obtained by reprobing the membranes with anti-α-tubulin antibody. In A, the results are representative of three independent experiments. In C, n = 4; *p < 0.05 vs. scramble. In E–K, n = 5; *p < 0.05 vs. scramble

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