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. 2015 Dec 17;10(12):e0145290. doi: 10.1371/journal.pone.0145290

Fig 1. Construction of primary ES cells and ESAQP5-shRNA recombinant lentiviral vectors.

Fig 1

After three cell divisions, the ES density was approximately 99%. Glandular epithelial cells were stained by cytokeratin 19 by immunocytochemistry (A, original magnification, ×400). Stromal cells were stained by Vimentin (B, original magnification, ×400). The bottom right corner of the black horizontal line is 10 μm. Flow cytometry showed that both CK19 and Vimentin positive cells accounted for 2.4% of the total, single CK19 positive accounted for 0.1%, single Vimentin positive accounted for 93.4%, and both CK19 and Vimentin negative accounted for 4.1% (C). The shRNA insert is 2155 bp, and the empty plasmid is 2155 bp (D). The sequencing showed that the cloned AQP5shRNA sequence was inserted correctly without mutation (E). pLKO-GFP-TRC–AQP5shRNA was transformed into ES cells and confirmed by GFP under the fluorescence field of view (F) and under the bright field view (G). The recombinant plasmids were selected (H). Con means the negative control of scrambled shRNA that inserted into pLKO-GFP after enzyme digestion.