Hypothalamic overexpression of NHLH2 prevents the development of experimental obesity. A, Transcript expression of Nhlh2 in the hypothalamus of mice fed an HFD for different periods of time. B, Transcript expression of Nhlh2 in the hypothalamus of fasting and refed mice. C, Protein levels of hypothalamic NHLH2 in mice acutely treated with leptin. D, Transcript expression of Nhlh2 obtained during the efficiency test of the lentiviruses infecting CLU189 cells with 10⁶ viral particles. E, Schematic representation of the protocols used for evaluation of the impact of modulating hypothalamic NHLH2 in mice submitted simultaneously to an HFD and lentivirus injections, the obesity prevention model. The transcript expressions of Nhlh2 (F,G), Pcsk1 (gene coding for PC1/3) (H,I), and Pomc (J,K) were determined in the hypothalamus of mice submitted to the protocol depicted in E. Mean daily food intake (L), body mass (M), weight gain (N), body adiposity (O,P), and epididymal white adipose tissue mean adipocyte size (Q,R) were determined in mice submitted to the protocol depicted in E. A, C, D, *p < 0.05 versus control (CTL). B, *p < 0.05 versus fast. F-O, Q, *p < 0.05 versus scrambled (SCc, scrambled fed chow; SCh, scrambled fed an HFD). ^#p < 0.05 versus overexpression (OEc, overexpression fed chow; OEh, overexpression fed an HFD). R, Red asterisk represents crown-like structures. Crossed lines indicate measurement of adipocyte areas. CT, Control; DXA, dual X-ray absorptiometry; eWAT, epididymal white adipose tissue; HYP, hypothalamus; KD, knockdown; KDc, knockdown fed chow; KDh, knockdown fed an HFD; OE, overexpression; OEc, overexpression fed chow; OEh, overexpression fed an HFD; SC, scrambled; SCc, scrambled fed chow; SCh, scrambled fed an HFD. A, F-N, n = 5-15; B-D, O-R, n = 5.