Figure 1
High glucose priming of IL-1β, stimulation of IL-1β on PDGF-BB production, and vice-versa. (a) Activation of the inflammasome NLRP3 and subsequent increase of pro-IL-1β protein levels were measured after 24 h stimulation of mesangial cells with Glc 30 mM. Osmotic control was Glc plus mannitol (Mtl), final concentration 30 mM. (b) 24 h stimulation of human mesangial cells with IL-1β 1 nM increased the levels of PDGFB mRNA. (c) In turn, a 24 h stimulation of mesangial cells with 25 ng/ml of PDGF-BB activated NLRP3 activity and increased pro-IL-1β protein levels. (d) Bio-Plex analysis of IL-1β, and PDGF-BB in media after high glucose, osmotic control, IL-1β and PDGF-BB 48 h stimulations. *P < 0.05. Data are reported as mean ± SEM. Samples under the LOD were considered equal to the lowest value in the measurable range (0.25 pg/ml for IL-1β and 13.96 pg/ml for PDGF-BB). (e) NLRP3 gene expression levels increased after 24 h stimulation with Glc 30 mM and PDGF-BB 25 ng/ml. Uncropped blots after chemiluminescence development and total protein stain free blots are presented in Supplementary Fig. S4a. Quantification of the bands in the western blot is reported in Supplementary Fig. S5.
