Mir 148a

Total RNA containing miRNAs was extracted using the mirVana miRNA Isolation Kit (Ambion) and reverse transcribed into cDNA using the miScript II RT Kit (Qiagen). qPCR was performed using the miScript PCR kit (Qiagen) on a ViiA 7 Real Time PCR System (Applied Biosystems, Grand Island, NY). Expression of miR-148a was normalized to RNU6-2_1 and was analyzed using the efficiency corrected ΔCt method. Primer assays; RNU6-2_1 and miR-148a (Qiagen).

Total RNA was extracted using RNeasy Mini Kit (Qiagen) and qRT–PCR was performed as described previously.^{8 (link)} Relative expression of each gene was normalized to 18S RNA. Primer sequences are listed in the Supplementary Table 3. For miRNA analysis, total RNA including small RNA was extracted using miRNeasy Kit (Qiagen). Pre-miRNA expression was analyzed as described previously.^{50 (link)} Mature miR-148a expression was detected using miScript SYBR Green PCR Kit (Qiagen) and probes for RNU6 (Qiagen) and miR-148a (Qiagen). The cDNA was synthesized using miScript II RT Kit (Qiagen) and was used to perform microarray analysis using a brain cancer miRNA PCR Array (SABiosciences, Frederick, MD, USA) according to the manufacturer's instructions. Array data were analyzed using miScript miRNA PCR array data analysis tools (Qiagen and SABiosciences).