RNA Extraction and Gene Expression Analysis

Total RNA from cell lines and tumors were extracted using RNeasy or miRNeasy Mini kits (Qiagen), respectively. CDNA were prepared as described earlier [47 (link)]. PCR using GoTaq Hot Start Kit (Promega) were performed with following primers: Ngfr-for 5'-GAATGCGAGGAGATCCCTGG-3', Ngrf-rev 5'-GGAGCAATAGACAGGAATGAGG-3', Snai1-for 5'-CACCCATACAGGTGAGAAGC-3', Snai1-rev 5'-TGTCCTGGATGACAGAACCA-3', Nefh-for 5'-GCAGCCAAAGTGAACACAGA-3, Nefh-rev 5'-CTGAATAGCGTCCTGGTAGG-3', Mash1-for 5'-TTGAACTCTATGGCGGGTTC-3', Mash1-rev 5'-GCCATCCTGCTTCCAAAGTC-3', human ALK-for 5'-TGTTGCCTCTCCTCGATGTG-3', human ALK-rev TGTCTTCTCCGCTAATGGTG-3', murine Alk-for 5'-TGCCAGAAGTGTGTTCAGAAC-3', murine Alk-rev 5'-CCCTTCCATGAAGGCTTCAG-3'. Primers for Snai2, Sox9, Sox10, Gfap, Gapdh were described [29 (link)]. Cycling reactions were 2min at 95°C followed by 35 cycles of 30s at 95°C, 30s at 60 to 65°C and 30s at 72°C, followed by 5min at 72°C.