Fig. 6.

Increased LCN-2 in human AMD and binding of NF-κB to the LCN-2 promoter. a In human samples, data from immunoblots show a significant increase in LCN-2 expression in early AMD (MGS2) compared to age-matched controls (MGS1), which persisted in the later stages of the disease (n = 5 control donors/and n = 3 donors/disease stage). b Immunofluorescence demonstrates LCN-2 expressing infiltrating cells (neutrophils stained with anti-neutrophil elastase) in the sub-macular choroid and retina of an early AMD patient (arrows). In age-matched control samples, many fewer neutrophils are detected (asterisk) and they are not positive for LCN-2. Bars = 50 μm. GCL ganglion cell layer, INL inner nuclear layer, ONL outer nuclear layer. c ChIP analysis of LCN-2 promoter-binding activity for NF-κB p65 subunit in retinal cells from Cryba1 KO mice (+/− LPS) showing association of NF-κB in the promoter region (− 3171) of the LCN-2 gene, but not in floxed controls. d Reverse ChIP analysis followed by western blotting indicated association between NF-κB and STAT1 in the same region as described in c. e Immunoblot shows significantly higher nuclear expression of NF-κB-p65 and p50 subunits in Cryba1 KO + LPS retinal cells, as compared to floxed control.

Reproduced with permission from Ghosh et al. [92]