Fig 2: miR-106a-5p targeting inhibits LKB1 expression. (A) Targeted binding site of miR-106a-5p and LKB1. Results of the dual luciferase reporter assay for (B) Calu-3 and (C) NCI-H661 cells. Semi-quantified protein expression levels of LKB1 in the (D) Calu-3 and (E) NCI-H661 cells transfected with miR-106a-5p mimic and NC. (F) Expression characteristics of LKB1 mRNA in lung adenocarcinoma tissues and adjacent normal tissues. (G) Relationship between miR-106a-5p and LKB1 mRNA. miR, microRNA; LKB1, liver kinase B1; NC, negative control; WT, wild-type; MUT, mutant' 3'UTR, 3'untranslated region. *P<0.05 vs. mimic-NC group or normal tissue.

Fig 3: The relative mRNA (A) and protein expression (B) of energy metabolism-related genes in the Longissimus thoracis muscles of the dams (n = 6) at 100 d of gestation. ACACA = acetyl-CoA carboxylase alpha; ACOX1 = acyl-CoA oxidase 1; CPT1A = carnitine palmitoyltransferase 1A; G6PDH = glucose-6-phosphate dehydrogenase; INSR = insulin receptor; NR1H3 = nuclear receptor subfamily 1 group H member 3; GR = glucocorticoid receptor; PCK1 = phosphoenolpyruvate carboxykinase 1; PCK2 = phosphoenolpyruvate carboxykinase 2, mitochondrial; PGC1A = peroxisome proliferator-activated receptor gamma coactivator 1 alpha; PRKAA2 = protein kinase AMP-activated catalytic subunit alpha 2; PRKAB1 = protein kinase AMP-activated noncatalytic subunit beta 1; STK11 = serine–threonine kinase 11; p-AMPKα = phospho-AMP-activated protein kinase; p-STK11 = phospho-STK11; GADPH = glyceraldehyde-3-phosphate dehydrogenase. ∗P < 0.05 and #0.05 ≤ P < 0.10 for the effect of diet. Dams in the control group were provided 100% of their nutrient requirements. Dams in the restricted group were provided 60% of the intake of the control group during 45 to 100 d of gestation and then realimented to 100% of nutrient requirements.

Fig 4: The relative expression of energy metabolism-related genes in the Longissimus thoracis muscles of the offspring. mRNA expression in the fetuses (A, n = 10) and in the kids (B, n = 8). Protein expression in the fetuses (C) and in the kids (D). ACACA = acetyl-CoA carboxylase alpha; ACOX1 = acyl-CoA oxidase 1; CPT1A = carnitine palmitoyltransferase 1A; G6PDH = glucose-6-phosphate dehydrogenase; INSR = insulin receptor; NR1H3 = nuclear receptor subfamily 1 group H member 3; GR = glucocorticoid receptor; PCK1 = phosphoenolpyruvate carboxykinase 1; PCK2 = phosphoenolpyruvate carboxykinase 2, mitochondrial; PGC1A = peroxisome proliferator-activated receptor gamma coactivator 1 alpha; PRKAA2 = protein kinase AMP-activated catalytic subunit alpha 2; PRKAB1 = protein kinase AMP-activated noncatalytic subunit beta 1; STK11 = serine–threonine kinase 11; p-AMPKa = phospho-AMP-activated protein kinase; p-STK11 = phospho-STK11; GADPH = glyceraldehyde-3-phosphate dehydrogenase. *P < 0.05 and #0.05 = P < 0.10 for the effect of diet; §P < 0.05 for the effect of sex; &P < 0.05 for the effect of litter size. Dams in the control group were provided 100% of their nutrient requirements. Dams in the restricted group were provided 60% of the intake of the control group during 45 to 100 d of gestation and then realimented to 100% of nutrient requirements.

Fig 5: miR-106a-5p/LKB1 exerts cancer-promoting effects through the AMPK pathway. Protein expression levels of (A) p-AMPK, p-TSC2 and p-mTOR in Calu-3 cells. Protein expression levels of (B) p-AMPK, p-TSC2 and p-mTOR in NCI-H661 cells. *P<0.05 vs. mimic-NC + vector-NC group; #P<0.05 vs. mimic + vector-NC group. TSC2, tuberin; miR, microRNA; NC, negative control; LKB1, liver kinase B1; AMPK, AMP-activated protein kinase; p-, phosphorylated.