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Fig. 7 | Parasites & Vectors

Fig. 7

From: Autophagic digestion of Leishmania major by host macrophages is associated with differential expression of BNIP3, CTSE, and the miRNAs miR-101c, miR-129, and miR-210

Fig. 7

Global analysis of differentially expressed mRNAs in L. m.-infected BMDM and MIF western blot analyses with protein extracts from L. m.-infected and HBSS-starved BMDM. Methods: (ae) BMDM from BALB/c mice were infected with L. m. promastigotes for 1 or 24 h. Uninfected control BMDM were incubated for the same amount of time in RPMI medium. Total RNA from 2 independent experiments was harvested from L. m.-infected BMDM or uninfected controls and hybridized with Affymetrix® chips. The BMDM were additionally incubated for 1 h in HBSS. Proteins were harvested and subjected to western blot analyses with specific antibodies against MIF. The western blots from 3 independent experiments were analyzed densitometrically. ACTB served as the internal loading control. Results: (a) Differentially expressed genes were detected in L. m.-infected BMDM 1 and 24 h p.i.. The results were presented in MA plots. Large dots represent probe sets, which had significant differential expression (FDR < 0.05). Dot colors correspond to the direction of gene expression changes (red dots = significant upregulation; blue dots = significant downregulation). b At 1 h p.i., 61 probe sets and 878 probe sets at 24 h p.i. had significant differential expression in L. m.-infected BMDM. 25 probe sets had significant differential expression at both time points. The results were presented in a Venn diagram. c MIF was overexpressed in L. m.-infected BMDM 24 h p.i. compared to uninfected control BMDM. d Results of densitometric analyses of the western blots and of the corresponding Affymetrix® chip analysis confirmed MIF overexpression at the mRNA and the protein level in L. m.-infected BMDM 24 h p.i. compared to uninfected control BMDM. e 28 autophagy-related genes in L. m.-infected BMDM 24 h p.i. were LISA members. The edges represent known protein-protein interactions determined in the AIN. The colors correspond to the direction of the gene expression changes (red = significant upregulation; blue = significant downregulation). L. m.-inf. = L. m.-infected, n.s. = not significant, p.i. = post infection, RFU = relative fluorescence units, * = genes of glycolysis, *** p ≤ 0.001

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