Journal: Scientific Reports
Article Title: C3a triggers formation of sub-retinal pigment epithelium deposits via the ubiquitin proteasome pathway
doi: 10.1038/s41598-018-28143-0
Figure Lengend Snippet: C3a causes specific deposition of Col IV and Col VI underneath the RPE that is prevented by C3aR antagonist. Immunolabeling with antibodies for ( a ) Col IV, ( c ) Col VI, ( e ) Col I, and ( g ) EFEMP1 of hfRPE cells treated with different doses of C3a or C3a + C3aR antagonist for 2 weeks. Images were taken with confocal. Orthogonal views show the basal deposition of the ECM proteins (left: apical, right: basal). Scale bars 50 µm. Average quantification of ( b ) Col IV, ( d ) Col VI, ( f ) Col I, and ( h ) EFEMP1 fluorescent signal of hfRPE cultures treated with different doses of rhC3a (C3aR-ant) or rhC3a plus C3aR antagonist (C3aR + ant) for 2 weeks. ( i ) Average quantification of immunostainings for ECM proteins after 4 weeks of treatment with C3a. (ANOVA. Data represented as mean ± SEM. n = 6/C3a dose *p < 0.05, **p < 0.01). ( j ) mRNA expression of COL4 and COL6 normalized to GAPDH after treatment with C3a for 2 weeks in the absence and presence (pattern bar) of C3aR antagonist (ANOVA. n = 3/−C3aR ant and n = 3/+ C3aR ant. Data represented as mean ± SD, **p < 0.01, ***p < 0.001, ****p < 0.0001).
Article Snippet: Primary antibodies used were: Col IV (AB6586, Abcam, Cambridge, MA), Col VI (AB6588), Col I (AB34710), FN (AB2413), EFEMP1 (SC33722, Santa Cruz Biotechnology, Santa Cruz, CA), and C3aR (HM2195, Hycult Biotech, Plymouth Meeting, PA).
Techniques: Immunolabeling, Expressing