10.1371/journal.pone.0023810.g003
Basha L. Stankovich
Basha
L. Stankovich
Esmeralda Aguayo
Esmeralda
Aguayo
Fatima Barragan
Fatima
Barragan
Aniket Sharma
Aniket
Sharma
Maria G. Pallavicini
Maria
G. Pallavicini
ESC with knockdown of junction molecules by shRNA maintain ability to differentiate to Flk-1 cells.
Public Library of Science
2011
knockdown
junction
molecules
shrna
differentiate
flk-1
2011-09-06 02:36:31
Figure
https://plos.figshare.com/articles/figure/_ESC_with_knockdown_of_junction_molecules_by_shRNA_maintain_ability_to_differentiate_to_Flk_1_cells_/409391
<p>A) shRNA constructs efficiently silence endogenous mRNA in mESC. Quantitative RT-PCR was performed on GFP expressing cells generated with lentiviral constructs containing shRNA specific to E-Cadherin, Cx43, ZO1 and ZO2 to analyze transcript expression level. Samples were normalized to GAPDH and expressed relative to transcript levels in untransfected D3-ESC (dashed line). Error bars represent standard deviation of n = 3 samples. B) Flk-1 expression does not significantly differ in knockdown ESC lines. ESC lines stably expressing shRNA sequences specific to E-Cad, Cx43, ZO-1 and ZO-2 and ESC lines with increased knockdown levels of E-Cad and ZO-1 were differentiated in EB for 5 days and Flk-1 expression was analyzed flow cytometrically using anti-Flk-1 conjugated to PE. Non-viable cells were excluded based on fluorescence of DAPI. Paired t-tests with D3-ESC were performed with n = 7 (Kd-E-cad, Kd-Cx43, Kd-ZO-1) or n = 6 (Kd-ZO-2). * p<0.05; *** p<0.0005. Error bars represent standard error of the mean.</p>