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. 2021 Feb 2;28(6):351–372. doi: 10.1038/s41434-021-00227-z

Fig. 4. Development of PITX3 MiniPromoters for corneal stroma cells.

Fig. 4

A Bioinformatic design of Ple253. ENCODE segments that potentially regulate the expression of the PITX3 gene are highlighted horizontally. Vertically highlighted genomic regions correspond to their color-matched segments included in the MiniPromoter design and are numbered as promoter (P). B Postnatal day 0 intravenous injection of Ple253-EmGFP, harvested 4 weeks later, led to robust expression in the stromal layer (Str) of the cornea, as determined from Hoechst staining and cell morphology. Scale bar, 100 µm. (For ubiquitous smCBA promoter see Fig. S1A). C Adult intrastromal injection of Ple253-EmGFP, harvested 4 weeks later, led to robust expression in the stromal layer, as determined by cell morphology. Scale bar, 50 µm. (For smCBA see Fig. S1E.) D Adult intrastromal injection of Ple253-EmGFP, harvested 4 weeks later, was quantified for epifluorescence intensity in stromal cells, determined by cell morphology, was and showed significantly lower (~1/5×) expression compared to smCBA (p < 0.001). (For smCBA see Fig. S1E.) EmGFP emerald green fluorescent protein, End endothelial layer, Epi epithelial layer, N number of cells counted, TF transcription factor, TSS transcription start site. Green, anti-GFP; blue, Hoechst.