2.3. Galectin-3 Knockdown

Stable Gal-3 knockdown was achieved by MSC or J774 macrophages by transduction with lentiviral vectors carrying a shRNA sequence targeting Lgals3 gene or scrambled control (Lgals3_shRNA1 5′-GATTTCAGGAGAGGGAATGAT-3′; one Lgals3_scrbl_shRNA 5′-AGGTATGAGTCGAGATTGAGA-3′), as previously described [18]. Culture medium was replaced and the cells were cultured for an additional 48 h, being assessed for GFP reporter gene expression by using an inverted fluorescence microscope (Eclipse Ti-E; Nikon, Tokyo, Japan). The cells were expanded and knockdown efficiency for each shRNA was evaluated by confocal microscopy and qPCR analyses.