FIGURE 2.

shRNA reduces expression of syndecan-1 but does not alter the myeloma cell phenotype. A, CAG myeloma cells were transduced with lentiviral vectors delivering control, syn1A, or syn1C shRNA sequences. Stable knockdown of syndecan-1 was confirmed by RT-PCR (top panel) and flow cytometry (bottom panel): IgG control antibody (shaded), control cells expressing wild-type levels of syndecan-1 (thin solid line), syn1A cells (dashed line), and syn1C cells (thick solid line). B, control cells (dashed line) and syn1A knockdown cells (solid line) were stained for markers of B cell lineage CD20, CD38, and CD45, or for syndecan-1, -2, or -4, or with control IgG antibody (shaded peak) and analyzed by flow cytometry. The gates were set on live, GFP+ cells. GAPDH, glyceraldehyde-3-phosphate dehydrogenase.