Fig. 8: Neutrophil gelatinase-associated lipocalin (NGAL) mediates pro-fibrotic effect of mineralocorticoid receptor (MR) activation.

a, b CCD-18Co cells silenced for NGAL with siNGAL were incubated with aldosterone (Aldo, 100 nM) with or without TGF-β (10 ng/mL) for 24 h (n = 4 from independent experiments). Relative protein levels of α-SMA (a), COL1 (b) and representative image of western blot (c) from CCD-18Co cells lysates (n = 4 per group; Two-sided Mann-Whitney test). d–f CCD-18Co cells were incubated with increasing concentration of recombinant hNGAL (5 ng/ml and 50 ng/ml; n = 5 from independent experiments). d Relative mRNA level of Acta2, Ccn2 and Col1a1 from CCD-18Co cells lysates (n = 5 per group; ordinary one way ANOVA with Tukey’s multiple comparisons test). e Representative and relative protein expression of α-SMA from CCD-18Co cells lysates (n = 5 per group; ordinary one way ANOVA with Tukey’s multiple comparisons test). f Representative gelatine zymography and quantification of MMP-2 activity from CCD-18Co cell lysates (n = 5 per group; ordinary one way ANOVA with Tukey’s multiple comparisons test). Data are presented as mean values +/− SEM.