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. Author manuscript; available in PMC: 2022 Jul 1.
Published in final edited form as: Cell Stem Cell. 2021 May 12;28(7):1248–1261.e8. doi: 10.1016/j.stem.2021.04.003

Figure 4: GPHA2 and IFITM3 marked qLSCs while IFITM3 supported the undifferentiated state in vitro.

Figure 4:

(A) Human cornea frozen sections were subjected to immunofluorescence staining of indicated markers. (B) Human limbal epithelial cells were transfected with endoribonuclease prepared small interfering RNA against GPHA2 (esiGPHA2) or control (esiCtl) and cells were immunostained for GPHA2. (C) Human limbal epithelial cells were cultured and maintained undifferentiated at low calcium (Day 0) or induced to differentiate (Day 7) and stained for the indicated markers. (D) High magnification of IFITM3 staining (day 0). (E-H) Undifferentiated cells were transfected with esiIFITM3 or esiCtl. (E-F) The expression of the indicated markers was examined 4–5 days post-transfection by quantitative real-time polymerase chain reaction (E) and immunofluorescent staining (F). (G-H) Transfectants were seeded at clonal density and allowed to grow for 2–3 weeks. Rhodamine-stained colonies are shown in (G) and quantification in (H). Nuclei were detected by DAPI counter staining (A-D, F). Scale bars are 50 μm or 10 μm (D). Statistical significance was calculated using t-test (*, p-value<0.05). Data are represented as mean ± SD.