Fig. 2. Complement competent human serum induces RPE atrophy in CFH(Y/Y)-iRPE and CFH(H/H)-iRPE.

a, b Transmission electron micrographs show disrupted tight junctions in CC-HS treated CFH(Y/Y)-iRPE2 as compared to CI-HS treated CFH(Y/Y)-iRPE2 (red arrowheads) (N = 4 biologically independent replicates, iRPE2, 4, 8, 9). c, d CC-HS treatment-induced stress fibers (arrowheads in d) and resulted in the loss of hexagonal morphology (F-ACTIN, yellow) in cells, compared to CI-HS treatment of CFH(H/H)-iRPE5 (N = 3 biologically independent replicates, iRPE5, 8, 9). e, f Transepithelial resistance (TER or Rt) (e) and phagocytic ability (f) of CFH(Y/Y)-iRPE and CFH(H/H)-iRPE under basal (CI-HS) and CC-HS treatment conditions (N = 7 biologically independent replicates iRPE1, 2, 3, 4, 5, 6, 7) for TER and phagocytosis. P values were determined by pairwise comparisons using two-tailed t test with 95% confidence interval and Bonferroni correction. The horizontal lines in the boxplots indicate the median, the boxes indicate the first and third quartiles, and the whiskers indicate 5th and 95th percentile. g–i Resting stage and physiological stimuli of apical 1 mM K + and 100 mM ATP induced transepithelial potential (TEP) of CI-HS and CC-HS-treated iRPE monolayers. Representative traces are shown in (g, h), quantification is shown in (i) (N = 10 independent experiments for CI-HS and N = 5 independent experiments for CC-HS, iRPE8). P values were determined by pairwise comparisons using two-tailed t test with 95% confidence interval and Bonferroni correction. The horizontal lines in the boxplots indicate the median, the boxes indicate the first and third quartiles, and the whiskers indicate 5th and 95th percentile. j, k Immunostaining for VIMENTIN (red) in CI-HS-treated CFH(Y/Y)-iRPE2 (j) and CC-HS-treated CFH(Y/Y)-iRPE1 (k). Arrowheads point to membrane staining in control cells that is missing in CC-HS treated iRPE (N = 6 biologically independent replicates, iRPE1, 3, 5, 6, 7, 8).