Fig. 2

Upregulation of Pik3r1-Foxo3a-cMyc pathway by miR-155 augments glucose metabolism. a, b qRT-PCR results of Foxo3a and cMyc genes in miR-155^ko/+ or miR-155^ko/ko cells. Data are normalized to mouse Rpl13a. Error bar means ± SEM (n = 3). *p < .0.05, **p < 0.01, ***p < 0.001. c Western blot results of FOXO3a and cMYC protein from miR-155^ko/+ or miR-155^ko/ko cells. β-ACTIN was used as loading control. d–i qRT-PCR results of Glut1 (d), Glut3 (e), Glut4 (f), Hk2 (g), Pkm2 (h), and Ldha (i) genes in miR-155^ko/ko cells after knockdown of Foxo3a by siRNA. (j) A graph showing qRT-PCR results of Pik3r1 gene in miR-155^ko/+ or miR-155^ko/ko cells. Normalized to mouse Rpl13a. Error bar means ± SEM (n = 3). *p < .0.05, **p < 0.01, ***p < 0.001. k Luciferase reporter activity for the Pik3r1 3′UTR. A Luciferase reporter vector containing Pik3r 3′UTR region or an empty vector was transfected into miR-155^ko/+ or miR-155^ko/ko cells and the luciferase activity was measured after 24 h. Error bar means ± SEM (n = 3). **p < 0.01, ***p < 0.001 (l). Western blot results of p85α, PI3K p100α, PI3K p110β, PDK1, p-AKT, AKT and cMYC proteins from miR-155^ko/+ or miR-155^ko/ko cells. β-ACTIN was used as a loading control. m–r qRT-PCR results of Glut1 (m), Glut3 (n), Glut4 (o), Hk2 (p), Pkm2 (q), and Ldha (r) genes after knockdown of the Pik3r1 in miR-155^ko/ko cells by siRNA. s A flow diagram presentation of LC-MS/MS results measuring glucose and its metabolites, from miR-155^ko/+ (Marked in black) or miR-155^ko/ko (marked in blue). miR-155^ko/+ cells with stable inhibition of miR-155 (miRZIP), were also analyzed and marked in red (miRZIP155_ko/+)