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FIGURE 2. The regeneration of LESCs at the limbus after innate stem cells ablation. (A) Immunostaining of potential LESCs markers (CK14, deltaN-p63, p75NTR, CD63, TSPAN7, IFITM3, <t>ATF3,</t> and ApoE), Cx43 and CECs marker CK12 in frozen sections of normal cornea. (B-D) Immunostaining of Cx43, ApoE, and CK12 in frozen sections of normal cornea and LESCs-ablation cornea at indicated days after the limbal epithelial removal. The limbus was shown. (E) The percentage of ApoE+ cells at the limbus, and MFI (limbal basal cells)/MFI (peripheral basal cells) of Cx43 and CK12 were determined to indicate the recovery of LESCs. N, normal cornea. Scale bars = 50 μm. Note that the limbus is localized based on the location of iris and ciliary body in the bright field and the expressions of LESC markers.
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FIGURE 2. The regeneration of LESCs at the limbus after innate stem cells ablation. (A) Immunostaining of potential LESCs markers (CK14, deltaN-p63, p75NTR, CD63, TSPAN7, IFITM3, <t>ATF3,</t> and ApoE), Cx43 and CECs marker CK12 in frozen sections of normal cornea. (B-D) Immunostaining of Cx43, ApoE, and CK12 in frozen sections of normal cornea and LESCs-ablation cornea at indicated days after the limbal epithelial removal. The limbus was shown. (E) The percentage of ApoE+ cells at the limbus, and MFI (limbal basal cells)/MFI (peripheral basal cells) of Cx43 and CK12 were determined to indicate the recovery of LESCs. N, normal cornea. Scale bars = 50 μm. Note that the limbus is localized based on the location of iris and ciliary body in the bright field and the expressions of LESC markers.
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FIGURE 2. The regeneration of LESCs at the limbus after innate stem cells ablation. (A) Immunostaining of potential LESCs markers (CK14, deltaN-p63, p75NTR, CD63, TSPAN7, IFITM3, <t>ATF3,</t> and ApoE), Cx43 and CECs marker CK12 in frozen sections of normal cornea. (B-D) Immunostaining of Cx43, ApoE, and CK12 in frozen sections of normal cornea and LESCs-ablation cornea at indicated days after the limbal epithelial removal. The limbus was shown. (E) The percentage of ApoE+ cells at the limbus, and MFI (limbal basal cells)/MFI (peripheral basal cells) of Cx43 and CK12 were determined to indicate the recovery of LESCs. N, normal cornea. Scale bars = 50 μm. Note that the limbus is localized based on the location of iris and ciliary body in the bright field and the expressions of LESC markers.
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FIGURE 2. The regeneration of LESCs at the limbus after innate stem cells ablation. (A) Immunostaining of potential LESCs markers (CK14, deltaN-p63, p75NTR, CD63, TSPAN7, IFITM3, ATF3, and ApoE), Cx43 and CECs marker CK12 in frozen sections of normal cornea. (B-D) Immunostaining of Cx43, ApoE, and CK12 in frozen sections of normal cornea and LESCs-ablation cornea at indicated days after the limbal epithelial removal. The limbus was shown. (E) The percentage of ApoE+ cells at the limbus, and MFI (limbal basal cells)/MFI (peripheral basal cells) of Cx43 and CK12 were determined to indicate the recovery of LESCs. N, normal cornea. Scale bars = 50 μm. Note that the limbus is localized based on the location of iris and ciliary body in the bright field and the expressions of LESC markers.

Journal: Investigative ophthalmology & visual science

Article Title: De-Differentiation of Corneal Epithelial Cells Into Functional Limbal Epithelial Stem Cells After the Ablation of Innate Stem Cells.

doi: 10.1167/iovs.65.13.32

Figure Lengend Snippet: FIGURE 2. The regeneration of LESCs at the limbus after innate stem cells ablation. (A) Immunostaining of potential LESCs markers (CK14, deltaN-p63, p75NTR, CD63, TSPAN7, IFITM3, ATF3, and ApoE), Cx43 and CECs marker CK12 in frozen sections of normal cornea. (B-D) Immunostaining of Cx43, ApoE, and CK12 in frozen sections of normal cornea and LESCs-ablation cornea at indicated days after the limbal epithelial removal. The limbus was shown. (E) The percentage of ApoE+ cells at the limbus, and MFI (limbal basal cells)/MFI (peripheral basal cells) of Cx43 and CK12 were determined to indicate the recovery of LESCs. N, normal cornea. Scale bars = 50 μm. Note that the limbus is localized based on the location of iris and ciliary body in the bright field and the expressions of LESC markers.

Article Snippet: The following primary antibodies were used: rabbit anti-CK12 monoclonal antibody (Abcam, ab185627; 1:400), rabbit anti-CK7 monoclonal antibody (Abcam, ab181598; 1:400), rabbit anti-connexin 43 (Cx43) monoclonal antibody (CST, #3512; 1:80), rabbit anti-ApoE monoclonal antibody (Abcam, ab183596; 1:400), rabbit anti-CK14 monoclonal antibody (Abcam, ab119695; 1:200), rabbit anti-deltaN-p63 polyclonal antibody (BioLegend, 619002; 1:400), rabbit anti-p75NTR monoclonal antibody (CST, #8238; 1:800), rabbit anti-CD63 polyclonal antibody (Bioworld, BS72936; 1:400), rabbit anti-TSPAN7 polyclonal antibody (Proteintech, 18695-1-AP; 1:100), rabbit anti-IFITM3 monoclonal antibody (CST, #59212; 1:100), mouse anti-ATF3 monoclonal antibody (Santa Cruz, sc-518032; 1:100), rabbit anti-YAP monoclonal antibody (CST, #14074; 1:100), rabbit anti-active-YAP (non-phosphorylated YAP) monoclonal antibody (Abcam, ab205270; 1:200), rabbit anti-Ki67 monoclonal antibody (CST, #9129; 1:100), rabbit anti-p-Histone H3-Ser10 (pH3) monoclonal antibody (CST, #53348; 1:100), and mouse antiCK15 monoclonal antibody (Santa Cruz, sc-47697; 1:200).

Techniques: Immunostaining, Marker