MiR-188 suppresses tumor initiation and progression in vivo . (A) CNE cells stably expressing miR-188 or miR-NC were injected into nude mice. MiR-188 (n =12) suppressed tumorigenesis compared with miR-NC (n =10). (B) miR-188 inhibits tumor growth in vivo. Tumor volumes were calculated by the length and width measured by vernier calipers every 2 days. (C) The weight of tumors from mice from (B). miR-NC (mean: 327.9 mg; range: 39.6 to 659.4 mg); miR-188 (mean: 83.2 mg; range: 19.5 to 136.6 mg). (D) Immunoblot analysis of cyclin D1, cyclin D3, cyclin E1, cyclin A2, CDK2, CDK4 and GAPDH in tumors treated with miR-NC or miR-188. (E) Relative protein expression was quantified by densitometric analysis, GAPDH was used as internal control (Student t test, *p < 0.05). (F) Immunoblot analysis of phosphor-Rb Ser811, phosphor-Rb Ser780 and total Rb in tumors treated with miR-NC or miR-188. (G) Relative Rb phosphorylation was quantified by densitometric analysis, total Rb was used as internal control (Student t test, *p < 0.05).