FIG 8.

LGALS8 is targeted by miR-579 and influences L. pneumophila replication. THP-1 cells were transfected with either the miR-579 mimic or the miRNA mimic pool (miR-125b, miR-221, and miR-579) at a final concentration of 30 nM. As a control, a scrambled precursor (scr) was transfected. At 24 h posttransfection, THP-1 cells were activated with PMA for another 24 h and then infected with L. pneumophila at an MOI of 0.5. (A) LGALS8 expression was examined with qPCR and is displayed as the log₂ fold change. Luciferase reporter assay analyses were performed in HEK-293T cells. The plasmid contained either the wild-type (LGALS8_WT) or the mutated (LAGLS8_mut) version of the 3′ UTR of LGALS8. (B) Ratios of Renilla and firefly luciferase luminescence were normalized to the vector without insert. BDMs were transfected with an siRNA pool targeting LGALS8 (siLGALS8) or with a scrambled siRNA as a control (scr) and infected at an MOI of 0.5 or left untreated as a control. (C) Downregulation of LGALS8 expression with siRNA was verified by qPCR and is displayed as the log₂-fold change. (D) CFU of Legionella were determined 24, 48, and 72 h postinfection after a knockdown of LGALS8. Boxes show the upper and lower quartiles, with medians (when n = 3 independent biological replicates) and whiskers indicating minimal and maximal values (when n = 4 or 5 independent biological replicates). Two-way ANOVA with Sidak’s correction was performed. *, P ≤ 0.05; **, P ≤ .01; ***, P ≤ 0.001; ****, P ≤ 0.0001 (compared to the corresponding control). #, P ≤ 0.05; ##, P ≤ 0.01; ###P ≤ 0.001 (compared to wild type) (B). ##, P ≤ 0.01 (for global treatment effects) (D).