Extended Data Fig. 2: BRP activates Fos expression and controls feeding without lowering blood glucose.
From: Prohormone cleavage prediction uncovers a non-incretin anti-obesity peptide
a, Peptide library design. 100 amidated peptides ranging from 5 to 25 amino acids length were synthesized with a C-terminal amide. b, Fos gene expression in NS-1 cells treated for 1 h with vehicle, NGF (50 ng/ml), BRP or scrambled BRP peptide (100 ug/ml). N = 3 biological replicates/group. c, Fos gene expression in Neuro2a cells treated with vehicle, or BRP peptide (50 µg/ml) for indicated times. N = 3 biological replicates/group. d, Fos gene expression in NS-1 cells treated with vehicle, or BRP peptide (50 µg/ml) for indicated times. N = 3 biological replicates/group. e, Fos gene expression in Neuro2a cells treated for 1 h with vehicle or BRP peptide every day for four days. N = 3 biological replicates/group. f, Fos gene expression in NS-1 cells treated for 1 h with vehicle, NGF (50 ng/ml), unmodified or amidated BRP (100 g/ml). N = 3 biological replicates/group. g, Fos gene expression in NS-1 cells treated for 24 h with vehicle or increasing doses of BRP peptide. N = 3 biological replicates/group. h, Fos gene expression in Neuro2a cells treated for 24 h with vehicle or increasing doses of BRP peptide. N = 3 biological replicates/group. i, Fos gene expression in Neuro2a cells treated for 1 h with vehicle or increasing doses of BRP peptide. N = 3 biological replicates/group. j, Fos gene expression in NS-1 cells treated for 1 h with vehicle or increasing doses of BRP peptide. N = 3 biological replicates/group. k, Fos gene expression in INS1 cells treated for 1 h with vehicle or increasing dose of GLP-1. N = 3 biological replicates/group. l, Pharmacokinetic levels of plasma and brain BRP after I.P. injection of 10 mg/kg BRP in lean C57BL/6 J male mice. N = 3 mice per group and per time point. m, Plasma levels of BRP, THRIL and LFNLC peptides after I.P. injection of 10 mg/kg BRP in lean C57BL/6 J male mice. N = 3 mice per group and per time point. n, Food intake measurement in lean C57BL/6 J male mice for up to 3 h after I.P. injection of vehicle, 5 mg/kg BRP or 5 mg/kg THRIL peptide. N = 3 independent experiments with 3 mice/group in each experiment. o, Food intake measurement in lean C57BL/6 J male mice for up to 3 h after I.P. injection of vehicle, 5 mg/kg BRP or 5 mg/kg LFNLC peptide. N = 3 independent experiments with 3 mice/group in each experiment. p, Food intake measurement in lean C57BL/6 J female mice for up to 3 h after I.P. injection of vehicle, 5 mg/kg BRP peptide, or 2 mg/kg GLP-1. N = 3 independent experiments with 3 mice/group in each experiment. q, Food intake measurement in C57BL/6 J DIO male mice for up to 3 h after I.P. injection of vehicle, 10 mg/kg BRP peptide, or 2 mg/kg GLP-1. N = 3 mice/group. r, Food intake measurement in lean C57BL/6 J male mice for up to 3 h after I.P. injection of vehicle or 5 mg/kg truncated BRP peptides. N = 3 independent experiments with 3 mice/group in each experiment. s, Fasting blood glucose levels in lean C57BL/6 J male mice for up to 3 h after I.P. injection of vehicle, 5 mg/kg peptides or 2 mg/kg GLP-1. N = 3 mice/group. t, Fasting insulin levels in lean C57BL/6 J male mice for up to 2 h after I.P. injection of vehicle or 5 mg/kg BRP. N = 3 mice/group. Data are presented as mean +/− S.E.M, P value = ns (non-significant), *P < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 by one way Anova (b, f) or two-way Anova for multiple comparisons (c, d, e, m, n, o, p, q, r, s).
