Fig. 7.

Lactadherin affects the whole proteome and transcriptome of MAECs.A, schematic representation of the experimental setup; 10 μg/ml or 500 ng/ml lactadherin was used for immobilized and soluble stimuli, respectively. Volcano plots highlight the up (red) and down (blue) significantly changed proteins (q-value <0.05 and absolute log2 fold change >0.58) after treatment with B, soluble lactadherin (Lact) and C, immobilized Lact for 24 h. Proteins not affected are shown in grey. Figure 7, B and C: The presence of lactaderin/Mfge8 likely indicates contamination by the recombinant protein used in the experimental procedure. D, heatmaps showing the proteins affected by soluble and immobilized lactadherin generated using SASP Atlas, http://www.saspatlas.com (38). The top 20 significantly affected proteins (ranked on q value) were listed in the SASP query tool, which displays a heatmap depicting the presence and secretion levels in different SASPs; human fibroblasts after induction of senescence by X-ray irradiation (Fibro IR), RAS overexpression (Fibro RAS) and Atazanivir treatment (Fibro ATV); human renal epithelial cells after induction of senescence by X-ray irradiation (Epi IR). E, GSEA shows the most affected pathways in MAECs cultured on immobilized lactadherin for 24 h using WebGestalt (64). F, bar plot shows relative gene expression quantified by RT-qPCR for different genes in MAECs. Data are shown as average ± SEM of fold change, normalized to control (No Lact) average transcript levels. n = 3; ∗∗p < 0.01; ∗p < 0.5, ordinary two-way ANOVA.