Research Paper Volume 11, Issue 6 pp 1648—1663

Figure 4. Effect of glycated albumin on FL-RAGE proteolysis in vitro. (A) Expression of FL-RAGE in R3/1-pLXSN (pLXSN) or R3/1-FL-RAGE (FL-RAGE) was determined using 20 µg of protein lysate with a specific antibody raised against the extracellular domain of RAGE. GAPDH detection was used as normalizer. (B) Quantification of cRAGE in the supernatant of R3/1-pLXSN (pLXSN) or R3/1-FL-RAGE (FL-RAGE) cells 6 h after medium changing by means of ELISA assay. Data are expressed as mean ± SD. ***, P≤0.0001. Unpaired t test ; n=4. (C) Quantification of cRAGE in the supernatant of R3/1-FL-RAGE cells treated or not (--) with indicated concentrations of control human albumin (HA) or glycated human albumin (Gly-HA) for 6 h by means of ELISA assay. Data are expressed as mean ± SD. **, P≤0.01. One-way ANOVA with Bonferroni post-hoc test; n = 4. (D) Representative Western blot (left panel) and corresponding quantification (right panel) of FL-RAGE expression using 5 μg protein lysates of R3/1-FL-RAGE cells treated or not (--) with 500 μg/mL HA or Gly-HA for 6 h. GAPDH detection was used as normalizer. Data are expressed as mean ± SD. *, P≤0.05. One-way ANOVA with Bonferroni post-hoc test; n=4; a.u., arbitrary units.