Journal: Journal of Translational Medicine

Article Title: Wnt signaling in age-related macular degeneration: human macular tissue and mouse model

doi: 10.1186/s12967-015-0683-x

Figure Lengend Snippet: Wnt signaling molecules in human retina. a Immunochemistry of phosphorylated LRP6 found higher activated LRP6 in the ganglion cell layer ( brown-blackish staining , arrows ) of the retinal sections of AMD maculae ( upper panel ) than in non-AMD macular tissues ( bottom panel ). Atrophic/degenerative photoreceptors and abnormal RPE cells are also noted in the macula of AMD specimens ( blue circles ). These areas also illustrate stronger immunoreactivities compared to the normal macula in the lower panel. NFL nerve fiber layer, GCL ganglion cell layer, IPL inner plexiform layer, INL inner nuclear layer, OPL outer plexiform layer, ONL outer nuclear layer. b mRNA expression of microdissected macular cells from human retinal section was measured by RT-PCR. Higher expression of CYCLIN D , cMYC , AXIN2 , and VEGF was observed in AMD patients compared to non-AMD controls (N = 5, Error bar mean ± SEM); * p < 0.05. c Immunochmistry of β-catenin in the macula. Higher expression of β-catenin was illustrated in AMD macular region compared to non-AMD macular area

Article Snippet: Polyclonal antihuman goat IgG antibodies against total LRP6 (Cell Signaling Technology, Danvers, MA) were applied at a 1:50 dilution for 60 min at room temperature.

Techniques: Staining, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Journal of Translational Medicine

Article Title: Wnt signaling in age-related macular degeneration: human macular tissue and mouse model

doi: 10.1186/s12967-015-0683-x

Figure Lengend Snippet: Western blot analysis of key components of the Wnt pathway in mouse retina. a Phosphorylated LRP6 and non-phosphorylated β-catenin were higher in the retina of Ccl2 − / − /Cx3cr1 − / − /rd8 compared to wildtype B6N mice. Phosphorylated LRP6 and non-phosphorylated β-catenin were measured in retinal homogenates of mice treated with the anti-LRP6 antibody or control mIgG. Reduced phosphorylated LRP6 and non-phosphorylated β-catenin were observed after the anti-LRP6 antibody treatment (n = 3 from each condition). b Quantitative analysis of 3 experiments, *: <0.05

Article Snippet: Polyclonal antihuman goat IgG antibodies against total LRP6 (Cell Signaling Technology, Danvers, MA) were applied at a 1:50 dilution for 60 min at room temperature.

Techniques: Western Blot

Journal: Journal of Translational Medicine

Article Title: Wnt signaling in age-related macular degeneration: human macular tissue and mouse model

doi: 10.1186/s12967-015-0683-x

Figure Lengend Snippet: Slower progression or alleviation of retinal lesions in the anti-LRP6 treated eyes as compared to the control in both models. a Representative fundoscopic images of Ccl2 − / − /Cx3cr1 − / − /rd8 . Arrows indicate the yellowish deep retinal lesions. b Comparison of the average retinal lesion scores between treated and untreated eyes of 35 Ccl2 − / − /Cx3cr1 − / − /rd8 mice. Error bar mean ± SEM; *: p < 0.01. c Representative fundoscopic images of Ccl2 − / − /Cx3cr1 gfp/gfp . Arrows indicate the yellowish deep retinal lesions. In Ccl2 − / − /Cx3cr1 gfp/gfp , only 5 out of 10 treated eyes but 7 out of 10 control eyes showed lesion progression

Article Snippet: Polyclonal antihuman goat IgG antibodies against total LRP6 (Cell Signaling Technology, Danvers, MA) were applied at a 1:50 dilution for 60 min at room temperature.

Techniques:

Journal: Journal of Translational Medicine

Article Title: Wnt signaling in age-related macular degeneration: human macular tissue and mouse model

doi: 10.1186/s12967-015-0683-x

Figure Lengend Snippet: Histology of the Ccl2 − / − /Cx3cr1 − / − /rd8 mouse retina. Representative histological micrographs: fewer lesions in anti-LRP6 treated eyes, as shown by the reduced loss of photoreceptors and more organized retinal structure. Black arrows indicate degenerative photoreceptors in the OPL ( top right panel ). Red arrows show degeneration and atrophy of ONL and loss of IS/OS in mIgG-injected group ( bottom right panel ). Some Crb1-associated lesions in the OPL are also seen ( asterisk , top left panel ) (H&E, original magnification, ×200). GCL ganglion cell layer, IPL inner plexiform layer, INL inner nuclear layer, OPL outer plexiform layer, ONL outer nuclear layer, IS / OS inner segments/outer segments of photoreceptors

Article Snippet: Polyclonal antihuman goat IgG antibodies against total LRP6 (Cell Signaling Technology, Danvers, MA) were applied at a 1:50 dilution for 60 min at room temperature.

Techniques: Injection

Journal: Journal of Translational Medicine

Article Title: Wnt signaling in age-related macular degeneration: human macular tissue and mouse model

doi: 10.1186/s12967-015-0683-x

Figure Lengend Snippet: Retinal A2E accumulation in the Ccl2 − / − /Cx3cr1 − / − /rd8 mice. Retinal A2E levels in anti-LRP6 treated and control eyes of Ccl2 − / − /Cx3cr1 − / − /rd8. A2E concentrations were determined by HPLC in the retina. The A2E level was reduced after anti-LRP6 antibody injection. n = 5 from each group; *: p < 0.05

Article Snippet: Polyclonal antihuman goat IgG antibodies against total LRP6 (Cell Signaling Technology, Danvers, MA) were applied at a 1:50 dilution for 60 min at room temperature.

Techniques: Injection

Journal: Journal of Translational Medicine

Article Title: Wnt signaling in age-related macular degeneration: human macular tissue and mouse model

doi: 10.1186/s12967-015-0683-x

Figure Lengend Snippet: mRNA expression of Wnt signaling pathway target genes in the treated and control eyes in Ccl2 − / − /Cx3cr1 − / − /rd8 mice. In general, there is lower transcript level in the eyes received anti-LRP6 compared to the control eyes measured by quantitative RT-PCR. *: p < 0.05

Article Snippet: Polyclonal antihuman goat IgG antibodies against total LRP6 (Cell Signaling Technology, Danvers, MA) were applied at a 1:50 dilution for 60 min at room temperature.

Techniques: Expressing, Quantitative RT-PCR