Sh rnas

To deplete endogenous CD26 mRNA, three small interfering (si) RNAs and two short hairpins (sh) RNAs were obtained from Qiagen (Hilden, Germany) or Sigma-Aldrich (reference sequence: NM_001935). The sequences are as follows.
CD26 siRNA-1: 5′-ACACTCTAACTGATTACTAA-3′,
CD26 siRNA-2: 5′-CAGTAAAGAGGCGAAGTATTA-3′CD26 siRNA-3: 5′-ATCGGGAAGTGGCGTGTTCAA-3′CD26 shRNA-1: 5′-CCGGGACTGAAGTTATACTCCTTAACTCGAGTTAAGGAGTATAACTTCAGTCTTTTTG-3′CD26 shRNA-2: 5′-CCGGCCAATGCAACTTCCATACAAACTCGAGTTTGTATGGAAGTTGCATTGGTTTTTG-3′To deplete endogenous CD9, two siRNAs and two shRNAs were used (reference sequence: NM_001769). The sequences are as follows.
CD9 siRNA-1: 5′-CGTGGAACAGTTTATCTCAT-3′CD9 siRNA-2: 5′-AATTGCCGTGGTCATGATATT-3′CD9 shRNA-1: 5′-CCGGGCTGTTCGGATTTAACTTCATCTCGAGATGAAGTTAAATCCGAACAGCTTTTTG-3′CD9 shRNA-2: 5′-CCGGCACAAGGATGAGGTGATTAAGCTCGAGCTTAATCACCTCATCCTTGTGTTTTTG-3′For controls, negative control siRNA (Qiagen) or non-target shRNA control (Sigma-Aldrich) was used.

Inducible reporter Mesp1/Gata4 and iGata cell lines were obtained from Professors Cedric Blanpain (Université Libre de Bruxelles, Belgium, (Bondue and Blanpain, 2010 (link))) and Todd Evans (Weill Cornell Medical College, USA, (Holtzinger et al., 2010 (link); Turbendian et al., 2013 (link))), respectively. Inducible Mesp1 cells were maintained and differentiated as previously described (Bondue and Blanpain, 2010 (link), Bondue et al., 2008 (link)). The iGata4/5/6 cells were maintained and differentiated as previously described (Holtzinger et al., 2010 (link), Turbendian et al., 2013 (link)), except that Embryoid Bodies were differentiated in hanging drop culture. shRNAs were obtained from Qiagen and stable transfected cell lines were generated according to manufacturer's instructions.