Ion Torrent Proton sequencing; RNA sequencing investigating the role of TNC in tumor progression using FVB TNC WT and TNC KO mice grafted with NT193 shC cells or NT193 shTNC cells
RNA sequencing investigating the role of TNC in tumor progression using FVB TNC WT and TNC KO mice grafted with NT193 shC cells or NT193 shTNC cells
NT193 cells transfected with control sh or shTNC The protocol is described in Sun et al., 2019 (PMID: 31288084). Briefly, silencing of Tnc  in mouse cells was done by short hairpin (sh) mediated gene expression knock down (KD). Lentiviral particles with shRNA vectors (Sigma- Aldrich) specific for Tnc  were used. sh1, TRCN0000312137, sequence 5' -CCGGCCCGGAACTGAATATGGGATTCTCGAGAATCCCATATTCAGTTCCGGGTTTTTG-3'; sh2, TRCN0000312138, 5'-CCGGGCATCAACACAACCAGTCTAACTCGAGTTAGACTGGTTGTGTTGATGCTTTTTG-30. Lentiviral particles encoding a non-targeting shRNA vector were used as a control (SHC202V, Sigma-Aldrich). Transduced cells were selected with normal medium supplemented with 10 m g/ml puromycin (ThermoFisher) and the selection pressure was maintained in all in vitro experiments. Whole RNA extraction from tissues Whole RNAseq