Figure 2.

Characterization of the SKO-B2M and DKO hESC-RPEs

(A) Immunofluorescence images of WT and SKO-B2M showing B2M, HLA-I, and ZO-1 expression. Magnified box for HLA-I shows dotted extracellular pattern in WT cells.

(B) Representative flow cytometry histogram showing the percentage of WT and SKO-B2M expressing extracellular HLA-I. Dotted line histogram shows HLA-I FMO (negative control used for gating).

(C) Western blot showing the HLA-I and B2M protein expression of WT and SKO-B2M cells.

(D) Gene expression analysis of HLA- and RPE-related genes in the targeted hESC-RPEs. Values are normalized to GAPDH and displayed as relative to WT cells.

(E) Immunofluorescence images of WT and DKO cells showing B2M, HLA-I, and ZO-1 expression. Magnified box for HLA-I shows dotted extracellular pattern in WT cells.

(F) Representative flow cytometry histogram showing the percentage of WT and DKO cells expressing extracellular HLA-I. Dotted line histogram shows HLA-I FMO (negative control used for gating).

(G) Gene expression analysis of pluripotent and HLA-related genes in the targeted hESC-RPEs. Values are normalized to GAPDH and displayed as relative to WT.

Bars represent mean ± SEM from three independent experiments.

Scale bars, 100 μm (A and E) and 50 μm zoom-in (A and E). Molecular weight of HLA-I = 43 kDa; B2M = 12 kDa. See also Figure S2.