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. 2017 Feb 28;114(11):E2243–E2252. doi: 10.1073/pnas.1614412114

Fig. S3.

Fig. S3.

Analysis of iOL transgene dependency using a tet-inducible lentiviral expression construct. (A) Tet-inducible SON was generated by cloning a polycistronic SON cassette into pHAGE, a third-generation lentiviral expression vector. An IRES-PAC cassette was introduced following the SON expression cassette allowing optional puromycin selection. (B) NPC were cotransduced with lentiviruses expressing tet-inducible SON and lentiviruses expressing rtTA. Expression of SON was induced by doxycycline for different time periods including 0 d as a negative control and 28 d as a positive control. Oligodendroglial lineage commitment was assessed by FACS analyses at day 28. Transduced NPC populations without doxycycline exposure (0) demonstrated comparable differentiation efficiencies as nontransduced NPC populations (Ctr), confirming the tight control of SON expression. Constitutive expression of SON (28 d) resulted in an increased differentiation efficiency compared with transient SON expression (7, 14, 21 d), indicating that a subpopulation of O4+ iOL was dependent on the ectopic expression of SON.