Fig. 3

Temporal LCN2 expression changes after retinal detachment for sequencing validation. (A) QRT-PCR analysis of Lcn2 mRNA levels in naïve and detached RPE at 1 dprd and 7 dprd (n = 6/group). (B) Representative immunoblot of LCN2 protein in RPE/choroid lysates from Lcn2 knockout mice (Lcn2^−/−), as well as RPE/choroid lysates from wildtype C57BL6/J mice under naïve, 1 dprd and 7 dprd conditions. Graph shows normalized results for 6 independent blots (n = 6 mice/group) showing mean ± SEM. Statistical analysis was performed using one-way ANOVA with repeated measures followed by Dunnett’s post hoc test. **p < 0.01. (C) IF co-localization of LCN2 (magenta) with Ezrin (green) and TMEM98 (red). Ezrin is an actin binding protein located in the apical microvilli of RPE cells, while TMEM98 is a transmembrane protein located at both the apical and basolateral surfaces of the RPE [46]. Ocular sections from C57BL6/J mice under naïve, 1 dprd and 7 dprd conditions and a section from Lcn2 KO mice with detached retina at 7 dprd. Staining for cell nuclei (blue) is shown in the merged images (right column) and magnified regions (boxes). Images were acquired using the Leica STELLARIS 8 FALCON Confocal Microscope, 40x objective