Fig. 4
From: Direct conversion of human fibroblast to hepatocytes using a single inducible polycistronic vector
Phenotype reversal of iHEP-LTDOX. a Schematic representation of time-course collection of reprogramming intermediates. Reprogramming of HDF-LTDOX was initiated at day 0 by addition of DOX 250 ng/mL. The collection point labeled 10+d represents cells reprogrammed for 10 days in DOX followed by specified days (in superscript) without DOX. b, c Expression of exogenous and endogenous reprogramming factors across a 0–17-day timeline. d, e Expression of exogenous transgenes and selected hepatic mRNA during the timeline described in a. mRNA levels were quantified by qRT-PCR and expressed relative to the expression at 10 days. f Scheme of the bicistronic lentiviral vectors independently expressing HNF4A, HNF1A, and FOXA3 together with mutually compatible fluorescent proteins. g HDF-LT cells were co-infected with HNF4A, HNF1A, and FOXA3 lentiviral vectors and cultured in HMM media. Expression of fluorescent proteins was assessed by flow cytometry after 30 days