Conversion of human fibroblasts into functional cardiomyocytes by small molecules by Nan Cao, Yu Huang, Jiashun Zheng, C. Ian Spencer, Yu Zhang, Ji-Dong Fu, Baoming Nie, Min Xie, Mingliang Zhang, Haixia Wang, Tianhua Ma, Tao Xu, Guilai Shi, Deepak Srivastava, and Sheng Ding Science Volume ():aaf1502 April 28, 2016 Published by AAAS

Fig. 1 Chemically induced CMs from HFFs. Chemically induced CMs from HFFs. (A) A representative contracting cluster induced by 15C at reprogramming day 30. Scale bars, 100 μm. (B to E) Number of contracting clusters (B to D) and flow-cytometry analyses of cTNT (E) in indicated conditions (n = 3). *P < 0.05 vs. 15C (B), 7C (C and D), and 9C (E). #P < 0.05 vs. +SU16F/JNJ. Nan Cao et al. Science 2016;science.aaf1502 Published by AAAS

Fig. 2 Structural and functional characterization of ciCMs. Structural and functional characterization of ciCMs. (A) Immunofluorescence analyses of CM markers. Insets show boxed areas at higher magnification. Scale bars, 25 μm. (B) Representative traces of ventricular-like APs and Ca2+ transients. Em, membrane potential. Dotted lines indicate 0 mV. F/F0, fluorescence relative to the baseline. Nan Cao et al. Science 2016;science.aaf1502 Published by AAAS

Fig. 3 ciCMs acquire transcriptional signatures of normal CMs. ciCMs acquire transcriptional signatures of normal CMs. (A) Hierarchical clustering analysis of genes that differently expressed in fibroblasts and fetal CMs, across all tested cell types. GO, gene ontology. (B) Cell/tissue classification heatmap of all tested cell types revealed by CellNet analyses. Higher classification scores indicate a higher probability that a query sample resembles the training sample. Nan Cao et al. Science 2016;science.aaf1502 Published by AAAS

Fig. 4 9C-treated fibroblasts convert into CMs in vivo. 9C-treated fibroblasts convert into CMs in vivo. Immunofluorescence analyses of heart sections after transplantation of control or 9C-treated HFFs. Insets show boxed areas at higher magnification. Scale bars, 100 μm. Nan Cao et al. Science 2016;science.aaf1502 Published by AAAS

Fig. 5 9C-treated fibroblasts have globally more open chromatin structure and increased chromosome accessibility on core cardiogenesis genes. 9C-treated fibroblasts have globally more open chromatin structure and increased chromosome accessibility on core cardiogenesis genes. (A) Heterochromatin foci shown by immunostaining for H3K9me3. Insets show a single cell nucleus at higher magnification. Scale bars, 100 μm. (B) Tracing of gene status in D6 and D11, relative to HFFs. Percentage within each category represents the ratio of genes show dynamic changes. Digits represent number of genes in each bar. (C) Levels of H3K4me3, H3K27ac, and H3K27me3 enrichment in genes in the GO term of “heart development.” Black/blue band inside the box represents the median/mean value, respectively. (D and E) ChIP analyses of β-catenin (D) or Smad1 (E) binding on promoters/enhancers of cardiogenic genes (n = 3). (F) A model for 9C-induced cardiac reprogramming. *P < 0.05; n.s, P > 0.05. Nan Cao et al. Science 2016;science.aaf1502 Published by AAAS