Journal: Scientific Reports

Article Title: Transcriptome and proteome characterization of surface ectoderm cells differentiated from human iPSCs

doi: 10.1038/srep32007

Figure Lengend Snippet: ( a ) Summarize the co-existence of non-neural ectoderm, neural ectoderm and neural ectoderm in mouse embryo Theiler stages (TS). Graphs show the markers the greatest differential expression in neural vs. non-neural (b ), neural vs. surface ectoderm (c) , embryo mesoderm vs. surface ectoderm (d ), surface ectoderm vs. embryo mesoderm ( e ), embryo endoderm vs. surface ectoderm (f ), and surface ectoderm vs. embryo endoderm (g ). Red stars: selected markers used in the following experiments.

Article Snippet: Primary antibodies are NANOG, TRA-1-81 (1:250, Stem cells); KRT8, KRT18, KRT19, AP-2γ, FOXG1 (1:200, Abcam); ALDH1A3, AP-2α, AFP (1: 200, Santa Cruz); Brachyury (T) (1:50, Novus); P63 (1:100, Genetex); p-SMAD1/5/9 (1:200, Cell Signaling); CDH1, Desmoplakin (1:500, BD Biosciences).

Techniques: Quantitative Proteomics

Journal: Scientific Reports

Article Title: Transcriptome and proteome characterization of surface ectoderm cells differentiated from human iPSCs

doi: 10.1038/srep32007

Figure Lengend Snippet: SE markers KRT8, KRT18, KRT19, p63, AP-2α, AP-2γ, ALDH1A3, p-SMAD1/5/9, CDH1, Desmoglein 3 and FOXG1 endodermal marker AFP, Mesodermal marker Brachyury (T), and neural ectoderm marker OTX2 were stained. DAPI was used to stain nuclei. The secondary antibodies Alexa −488 and Alexa −594 were used to visualize green and red signals, respectively. Insert in AFP staining image (+): Hela control cells stained by anti-AFP antibody and showed positive staining. Original magnification x200. Bars: 50 μm.

Article Snippet: Primary antibodies are NANOG, TRA-1-81 (1:250, Stem cells); KRT8, KRT18, KRT19, AP-2γ, FOXG1 (1:200, Abcam); ALDH1A3, AP-2α, AFP (1: 200, Santa Cruz); Brachyury (T) (1:50, Novus); P63 (1:100, Genetex); p-SMAD1/5/9 (1:200, Cell Signaling); CDH1, Desmoplakin (1:500, BD Biosciences).

Techniques: Marker, Staining, Control

Journal: Scientific Reports

Article Title: Transcriptome and proteome characterization of surface ectoderm cells differentiated from human iPSCs

doi: 10.1038/srep32007

Figure Lengend Snippet: ( a ) Cell morphologies in hiPSC (control), SE induction (SE, using BMP4 and DAPT), and SE induction with TGFβ-RI inhibitor SB431542 (two separate experiments named as SE + SB-1 and SE + SB-2) groups. Bar: 100 μm. Original magnification x100. ( b ) Immunoblotting analysis showed the expression of SE and other lineage markers in all groups. Actin was used as loading control. SE + SB-1 and SE + SB-2 suggested two separate experiments. Red stars: greatly altered compared to SE. Black box: results from SE group. Dashed black box: results from SE + SB groups. The gels have been run under the same experimental conditions. Full-length blots are shown in . (c ) Relative intensity is represented by selected marker expression in SE+SB groups compared to SE only group and plotted. Plotted data represent three independent experiments. *p < 0.05 (relative to SE groups). (d ) Modified protocol for in vitro differentiation of hiPSCs to SE. We modified the existing protocol by adding TGFβ-RI signaling inhibitor to improve the SE differentiation.

Article Snippet: Primary antibodies are NANOG, TRA-1-81 (1:250, Stem cells); KRT8, KRT18, KRT19, AP-2γ, FOXG1 (1:200, Abcam); ALDH1A3, AP-2α, AFP (1: 200, Santa Cruz); Brachyury (T) (1:50, Novus); P63 (1:100, Genetex); p-SMAD1/5/9 (1:200, Cell Signaling); CDH1, Desmoplakin (1:500, BD Biosciences).

Techniques: Control, Western Blot, Expressing, Marker, Modification, In Vitro