International Journal of Stem Cells : eISSN 2005-5447

Fig. 2.

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Fig. 2. mRNA and protein expression of RPE markers. (A) The mRNA expression of RPE-65, BEST, CRLBP, MITF, PEDF, and ZO-1 was evaluated by quantitative real-time polymerase chain reaction. mRNAs were quantified from hES-RPE, ARPE-19, fetal RPE, and adult RPE normalized to the geometric mean of a housekeeping gene (18s rRNA). (B) The mRNA expression of pluripotency (OCT4 and NANOG), neuroectoderm (PAX6) and RPE markers (MITF, CRLBP, BEST, RPE-65, PEDF and ZO-1) was also compared between undifferentiated hESCs and hESC-RPE. As expected, compared to undifferentiated H1, differentiated cells at passage 2 presented lower expression of pluripotency genes, and higher expression of neuroectoderm and RPE differentiation markers. (C) Protein expression of BEST and MITF was evaluated in p2 hESC-RPE cells by immunofluorescence. As expected, BEST presented membrane localization, and MITF, nuclear localization. (D) Western Blotting analysis of RPE-65 and CRLBP expression by p1, p2 and p3 hESC-RPE cells. Similar to fetal RPE, hESC-RPE cells from all the analysed passages presented detectable expression of both RPE markers. Bars represent standard error of the mean. Abbreviations: hES-RPE, RPE derived from human embryonic stem cells; fRPE, fetal RPE.
International Journal of Stem Cells 2021;14:74-84
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