Validation of the expression of novel miRNAs. A. Summary of the RT-PCR analysis of the 11 selected novel miRNAs. Novel candidates with extremely lower abundance can be detected by deep-sequencing but not by PCR amplification. Counts mean the numbers of each unique read coming from deep sequencing. B. Validation of the expression profile of 2 highly expressed novel miRNA candidates (candidate 11, candidate 55) by qPCR. All experiments were repeated for 3 times. Error bars represent standard deviation. The relative expression was normalized to E10 value for both qPCR and TPM. There is a high correlation between the deep-sequencing and qPCR results. C. Expression of the candidate in different tissues. The relative expression of miRNAs in different tissues was normalized to that of cortex. Note that all of them are enriched in the central nerve system. Experiments were repeated for 4 times. Error bars represent standard error deviation (n = 4; *P < 0.05). D. Regulation of C6 cell proliferation by Candidate 11. Overexpression of Candidate 11 promoted cell proliferation, whereas suppressing the endogenous Candidate 11 by overexpression of a sponge RNA (6X) reduced cell proliferation. Experiments were repeated for 3 times. Error bars represent standard error of the mean (n = 3; *P < 0.05).