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Fig. 2 | BMC Cancer

Fig. 2

From: MiR-214-3p targets Ras-related protein 14 (RAB14) to inhibit cellular migration and invasion in esophageal Cancer cells

Fig. 2

Effects of modulation of miR-214-3p on RAB14 protein and mRNA expression levels in human esophageal cells. (A) Levels of miR-214-3p in TE7, FLO-1 and SK-GT-4 cells transfected with pre-miR-214-3p (50 nM) as measured by q-PCR. (B) Changes in RAB14 protein expression after overexpressing miR-214-3p (50 nM) in TE7, FLO-1 and SK-GT-4 cells. Protein loading was assessed by GAPDH. The blots in this figure were cut prior to hybridization with antibody to RAB14 and GAPDH. Relative signal intensity was calculated as explained in Fig. 1E. (C) Levels of miR-214-3p after transfection of anti-miR-214-3p (25 nM) for 48 hrs in hESO cells as measured by q-PCR. Small nuclear U6 RNA was used as a control. Error bars represent mean ± S.D. * represents p < 0.05. (D) Effect of miR-214-3p silencing on RAB14 protein expression in hESO cells. hESO cells were transfected with anti-miR-214-3p (25 nM) for 48 hrs. Immunoblot was performed for RAB14 protein expression and GAPDH was used as a loading control. The blot in this figure was cut prior to hybridization with antibody to RAB14 and GAPDH. Relative signal intensity was calculated as explained in Fig. 1E. (E) Changes in levels of RAB14 mRNA in TE7, FLO-1 and SK-GT-4 cells following transfection of pre-miR-214-3p (50 nM). (F) Levels of RAB14 mRNA in hESO cells after transfection of anti-miR-214-3p (25 nM). Levels of RAB14 mRNA were measured by q-PCR. GAPDH was concurrently amplified to serve as an internal control. Error bars represent mean ± S.D. * signifies statistical significance (p < 0.05). Stability of RAB14 mRNA in (G) FLO-1 cells following transfection of pre-miR-214-3p and in (H) hESO cells after silencing miR-214-3p. Total RNA was isolated at indicated time points after administration of Actinomycin D (4 μM) and the remaining levels of RAB14 mRNA were measured by q-PCR. Levels were normalized with GAPDH. The half-life was calculated from the first order equation t1/2 = ln2/k

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