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Fig. 4 | Molecular Cancer

Fig. 4

From: Circular RNA circNRIP1 acts as a microRNA-149-5p sponge to promote gastric cancer progression via the AKT1/mTOR pathway

Fig. 4

Knockdown of circNRIP1 reduces the expression of AKT1. a. We performed a dual-luciferase reporter assay to determine the direct binding between 149-5p and AKT1 in GC cells. b. We found that AKT1 was significantly upregulated in GC tissues (415 GC tissues vs 34 normal tissues), and patients with high levels of AKT1 (298 GC tissues vs 94 normal tissues) had lower OS based on an analysis of the TCGA database. c, d. We subsequently detected higher levels of AKT1 in GC tissues than in adjacent normal tissues by performing immunochemistry analysis on 12 paired GC tumour tissues and normal stomach tissues, scale bar = 200 μm. e. qRT-PCR further confirmed the expression level of AKT1 and showed that it was upregulated in 80 GC tissues relative to adjacent normal tissues. f. We detected the circNRIP1 level in both groups and determined that the level of circNRIP1 in the high AKT1 group(n = 40) was significantly higher than that in the low AKT1 group (n = 40). g. Two circNRIP1 small interfering RNAs (siRNAs) specifically targeting the backsplice junction sequences at different binding sites in circNRIP1 were designed. h. Immunofluorescence assay showed that circNRIP1 knockdown successfully inhibited the expression level of AKT1, scale bar = 50 μm. All data are presented as the mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001

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