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. 2019 Sep;189(9):1814–1830. doi: 10.1016/j.ajpath.2019.05.022

Figure 3.

Figure 3

LS-HepcKO mice have labile iron accumulation in the neurosensory retina (NSR). A: Graphs of relative transferrin receptor (Tfrc) mRNA levels, determined by real-time quantitative PCR (qPCR), in NSR of LS-HepcKO mice versus controls (Hepc+/+;Alb-Cre+) at the indicated ages. Groups marked with a different symbol are significantly different from each other (P < 0.05), as determined by two-way analysis of variance with post hoc Tukey adjustment. B: Western blot analysis showing Tfr protein levels in the NSR in the LS-HepcKO mice versus controls at 12 months and relative pixel density of Western blot analysis Tfr bands corrected for glyceraldehyde-3-phosphate dehydrogenase (GAPDH). C: Relative Dmt1 mRNA levels in NSR of LS-HepcKO mice versus controls (Hepc+/+;Alb-Cre+ or Hepcflox/+;Alb-Cre+). D–O: Ferritin-L (Ft-L) immunolabeling of retinal sections. D, F, H, J, L, and N: At all three ages (3, 6, and 12 months), there is increased Ft-L immunolabeling in the NSR of LS-HepcKO mice (F, J, and N) compared with age-matched controls (D, H, and L). P: Pixel density of Ft-L immunolabeling in the NSR was quantified. Q: Relative levels of Hepc mRNA in LS-HepcKO NSR versus controls, determined by qPCR at 6 months. R and S: Inductively coupled mass spectrometry of NSR from LS-HepcKO and control mice at 6 months. R: There is no difference in iron (ppm) within the NSR of the LS-HepcKO mice when compared with controls. S: There is no change in the iron/zinc ratio in the NSR of LS-HepcKO mice compared with controls. Data are expressed as means ± SEM (A–C and P–S). ∗P < 0.05, ∗∗P < 0.01 (B and Q). Scale bars = 25 μm (D–O). GCL, ganglion cell layer; INL, inner nuclear layer; IPL, inner plexiform layer; ONL, outer nuclear layer; OPL, outer plexiform layer; RPE, retinal pigment epithelium.