Transfection of miR-150 into T-cell lymphoma cell lines reduces invasion and metastasis. (A) Visceral invasion of CTCL cells in NOG mice. Photographs show invasion/metastasis into visceral organs by hematoxylin and eosin and GFP staining of liver (original magnification ×200) and kidney (original magnification ×100), GFP staining of lung (original magnification ×200) in GFP-empty (control) transduced My-La (shown as “GFP-My-La”), and HH (shown as “GFP-HH”) cells. Invading CTCL cells or the invaded region is surrounded by broken lines. (B) Invasion/metastasis by transfected My-La and HH (GFP-empty [control] or GFP-miR-150) in NOG mice. Photographs show spleen and BM from NOG mice inoculated with GFP-empty (control) (shown as “GFP-control”), or GFP-miR-150-CTCL cells. (C) Flow cytometric analysis of BM and spleen invasion by My-La and HH transfectants (GFP-empty control [n = 10 each] or GFP-miR-150 [n = 10 each]). Bars are means ± SEM of 3 independent experiments. Asterisks (*) indicate statistical significance: **.001 ≤ P < .01; ***P < .001. (D) Cell migration assay. Shown are RFUs (Ex 485/Em 538) from ATN-1, MJ, and HUT78 cells transfected with GFP-empty (control) (shown as “control”) or GFP-miR-150, and My-La and HH cells transfected with GFP-miR-16-1, GFP-miR-29a, or GFP-miR-150. Migration was stimulated by 2% HS in the lower chamber; no serum was added to the upper chamber. Incubation time was 16 hours and cell Lysis buffer were transferred to a 96-well plate, and fluorescence was measured by plate reader at 480 nm/520 nm. P values were calculated using Student t test. Asterisks (*) indicate statistical significance: ***P < .001. Bars are means ± SEM of 3 independent experiments. (E) A schematic illustration of the migration assay is shown besides the bar graphs. NS, not significant.