Figure 4

MGO upregulates cell cycle genes and downregulates genes related to ECM organisation in GECs co-cultured with podocytes. (A) Volcano plot of gene expression changes in GECs exposed to 96 h of MGO compared to control cells. Genes with log₂FC > 1 and FDR-corrected p-values < 0.05 were marked in red (strongly upregulated genes); genes with log₂FC < − 1 and FDR-corrected p-values < 0.05 were highlighted in blue (strongly downregulated genes). (B) Heatmaps of the top 20 up- and downregulated genes in GECs after 96 h of MGO. (C) Functional annotation showing overrepresented gene ontology (GO) biological processes for the upregulated (red bars) and downregulated (blue bars) genes in GECs after 96 h of HG. Abbreviations used: neg., negative; pos., positive; reg., regulation; reg. of G1/S transition of mitotic cell cycle, regulation of transcription involved in G1/S transition of mitotic cell cycle. (D,E) Interaction network of the MGO-regulated genes belonging to the top GO terms in GECs, namely “G1/S transition of mitotic cell cycle” for the upregulated genes (D) and “extracellular matrix organization” for the downregulated genes (E), generated using the STRING database, based on high-confidence interactions (interaction score ≥ 0.7). (F,G) qPCR analysis of the expression of selected candidate genes that were among the prominent upregulated genes (CYP1A1, ID1, ID3, CYGB, IL24, red bars) and the prominent downregulated genes (FN1, FBN1, blue bars) in GECs after 96 h of MGO, based on the RNA-seq results. The relative expression levels of the genes in co-cultures of GECs with podocytes (F) and in GEC monocultures (G) are shown. For both co-cultures and monocultures n = 6.