Distribution of single nucleotide polymorphisms rs10490924 in <i>ARMS2</i> and rs1061170 in <i>CFH</i> and trend test of AMD severity stages (data of other SNPs is shown in S1 Table).

<p>Distribution of single nucleotide polymorphisms rs10490924 in <i>ARMS2</i> and rs1061170 in <i>CFH</i> and trend test of AMD severity stages (data of other SNPs is shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0156778#pone.0156778.s001" target="_blank">S1 Table</a>).</p

Univariate regression analysis for SNPs rs10490924 in ARMS2 and rs1061170 in CFH in patients with unilateral nAMD and different severity stages of the fellow-eye.

<p>Univariate regression analysis for SNPs rs10490924 in ARMS2 and rs1061170 in CFH in patients with unilateral nAMD and different severity stages of the fellow-eye.</p

Multivariate regression model for each severity stage with results for SNPs rs10490924 in <i>ARMS2</i> and rs1061170 in <i>CFH</i> (data of other SNPs is shown in S3 Table).

<p>Multivariate regression model for each severity stage with results for SNPs rs10490924 in <i>ARMS2</i> and rs1061170 in <i>CFH</i> (data of other SNPs is shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0156778#pone.0156778.s003" target="_blank">S3 Table</a>).</p

Grading results.

<p>Grading results.</p

Demographics.

<p>Demographics.</p

Area under the curve (AUC) of risk models in different AMD severity stages.

<p>Area under the curve (AUC) of risk models in different AMD severity stages.</p

Characteristics of the study subjects.

Characteristics of the study subjects.</p

Measurement of sphingolipid species in serum of late stage AMD patients with GA or CNV compared to healthy controls.

Measurement of sphingolipid species in serum of late stage AMD patients with GA or CNV compared to healthy controls.</p

Alteration of the sphingolipid metabolism under MAA-BSA treatment in WERI-Rb1 cells.

<p><b>(A)</b> The sphingolipid pathway depicting the gene symbols of the enzymes involved (complete protein names in <b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0200739#pone.0200739.s002" target="_blank">S2 Table</a></b>). <b>(B-H)</b> Cells were treated with 80 μg/mL of BSA or MAA-BSA for 24 h, and gene expression or lipid levels were measured. <b>(B)</b> Genes from the <i>de novo</i> and salvage ceramide synthesis pathways. <b>(C)</b> Genes involved in the synthesis of SM from ceramides, and synthesis of glucosylceramides (GlcCer) and galactosylceramides (GalCer), both hexosylceramides (HexCer). <b>(D)</b> Genes involved in the generation of ceramide by sphingomyelin (SM) hydrolysis. <b>(E)</b> Ceramide (Cer) species levels. <b>(F)</b> SM species levels; only results from most abundant species are shown. <b>(G)</b> Ratio of total ceramide to total SM levels. <b>(H)</b> Ratio of total ceramide to total HexCer levels. Each value corresponds to the mean ± SEM of six-fold independently performed replicates evaluated in two batches. Statistics: Linear regression adjusted for batch of analysis; ***p<0.001, **p<0.01, *p<0.05.</p

Influence of FHL-1 variants in expression of ceramide metabolism genes.

(A) Schematic representation of CFH and FHL-1 proteins. CFH is composed of 20 short consensus repeats (SCR), and its alternative splicing variant FHL-1 of 7 SCR and a unique extension of 4 amino acids. The complement regulatory region (SCR 1–4), and surface recognition domains (SCR 19–20) are indicated. Residue 402 is located in the SCR 7 of both proteins. (B-G) WERI-Rb1 cells treated with 80 μg/mL BSA or MAA-BSA during 24 h with or without addition of 200 μg/mL CFH or 80 μg/mL FHL-1 proteins. Expression of genes from (B) NAD(P)H dehydrogenase [quinone] 1 (NQO1), (C) Phosphatidylcholine:ceramide cholinephosphotransferase 1 (SGMS1), (D) Serine palmitoyltransferase 1 (SPTLC1), (E) Sphingolipid delta(4)-desaturase DES1 (DEGS1), (F) Ceramide synthase 2 (CERS2), (G) Ceramide glucosyltransferase (UGCG). Each value corresponds to the mean ± SEM of four-fold independently performed experiments. Statistics: two-way blocked ANOVA and Tukey’s test (no significant interaction between principal effects was observed); (C-G) only the significance from the Tukey’s test comparing PBS, CFH, FHL-1:Y402 (Y), and FHL-1:H402 (H) are shown in the graphs; all genes showed significant differences between MAA-BSA and BSA treatments: (C, F) p<0.001, (G, E) p<0.01, (D) p<0.05. ***p<0.001, *p<0.05.</p