Glut1 siRNA and shRNA Plasmids (h)

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	UM-UC-3 and HT-1376 bladder cancer cells were transfected in 6- or 96-well culture plates, at 60–80% confluence, with galectin-1 and GLUT1, respectively. Cells were also transfected with a scrambled siRNA in parallel as controls.

Protocol tips
UM-UC-3 and HT-1376 bladder cancer cells were transfected in 6- or 96-well culture plates, at 60–80% confluence, with galectin-1 and GLUT1, respectively. Cells were also transfected with a scrambled siRNA in parallel as controls.

Publication protocol

Galectin-1 or GLUT1 was depleted in human bladder cancer cells using a pool of three target-specific 20–25 nt siRNA (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA). UM-UC-3 and HT-1376 bladder cancer cells were transfected in 6- or 96-well culture plates, at 60–80% confluence, with galectin-1 and GLUT1, respectively. Cells were also transfected with a scrambled siRNA in parallel as controls.

For each transfection, cells were treated for 5 h with 2.4 µM of siRNA in transfection medium (Santa Cruz) containing 0.5 µL/cm2 of transfection reagent (Santa Cruz). After incubation, complete media was added and the cells were incubated for 24 or 48 h. Galectin-1 or GLUT1 downregulation was evaluated 24 h or 48 h post-transfection by Western blotting. The uptake and PDT experiments were performed 24 h or 48 h post-transfection with GLUT1 hsiRNA or galectin-1 hsiRNA, respectively.

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