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. 2010 Sep 20;20(6):1021–1029. doi: 10.1089/scd.2010.0198

FIG. 3.

FIG. 3.

Knockdown of SDF-1 secretion in CD133dMSCs by lentiviral short hairpin RNA (shRNA). CD133dMSCs were transduced with puromycin-selectable lentivectors expressing GFP (transduction control), scrambled (nonspecific) shRNA, or sequence-specific shRNA (against SDF-1). (A) Fluorescent activated cell sorting (FACS) of control cells (no label) and those transduced with GFP lentivector and selected by puromycin to determine cell purity after selection regime. (B) Enzyme-linked immunosorbent assay for SDF-1 showed a 94% and 89% reduction in level of secreted SDF-1 from CD133dMSCs after lentiviral transduction of 2 different shRNA sequences for the SDF-1 transcript compared with a scrambled vector. SDF-1 secretion levels were not different from cells that received the scrambled vector and cells that were not transduced (no vector control). All measurements were performed in triplicate. (C) Phase-contrast microscopy images of CD133dMSCs that received lentivectors and no vector control cells (magnification 10×). shSDF-1, shRNA specific to SDF-1; shScram, scrambled non-specific shRNA (control).