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Fig. 3 | BMC Cancer

Fig. 3

From: Malignant canine mammary epithelial cells shed exosomes containing differentially expressed microRNA that regulate oncogenic networks

Fig. 3

qRT-PCR validation of RNAseq data. a-c Relative quantification (log10) for selected validation targets miR-18a (a), miR-19a (b), and miR-181a (c). Relative quantification was calculated for each biological replicate according to the eq. 2-ΔCq, with cel-miR-39 as spike-in exogenous control and miR-16 as an endogenous control; experiments were performed in triplicate from cell culture to RNA extraction, cDNA synthesis and qRT-PCR. Data were not normally distributed and compared by non-parametric, non-directional Mann-Whitney test. p < 0.05 was considered statistically significant. The black horizontal line represents the group mean and the vertical “whiskers” represent ±1 SD. d Comparison of fold-change between microRNA deep-sequencing and manual stem-loop qRT-PCR assays for selected targets miR-18a, miR-19a, and miR-181a. Data were normalized using the 2-ΔΔCq method. The average group Cq for cfa-miR-16 (endogenous control) and cel-miR-39 (exogenous spike-in control) were used as housekeeping genes for normalization. White bars represent relative fold-change for RNAseq data, black bars represent fold-change for qRT-PCR (3 experimental replicates)

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