Journal: Cancer research communications
Article Title: STAG2 expression is associated with adverse survival outcomes and regulates cell phenotype in muscle-invasive bladder cancer
doi: 10.1158/2767-9764.crc-22-0155
Figure Lengend Snippet: A. Western blot demonstrating STAG2 protein expression in T24 cells transfected with scrambled control or STAG2 siRNA for 24, 48, and 72 hours. GAPDH used as loading control. Figure is representative of three independent experiments. B. T24 cells transfected with scrambled control or STAG2 siRNA, were seeded on top of a Transwell insert and allowed to travel through Matrigel-coated membrane (top panels, invasion) or uncoated membrane (bottom panels, migration) for 24 hours. C. Average number of T24 control or siSTAG2 cells per field view that migrated through uncoated membrane (left panel) or invaded through Matrigel-coated membrane (right panel) after 24 hours. D. T24 cells treated with scrambled control, shSTAG2-1, or shSTAG2-2, seeded in the top of a Transwell insert and allowed to move through Matrigel-coated membrane (top, invasion) or uncoated membrane (bottom, migration) for 24 hours. E. Average number of T24 control, shSTAG2-1, or shSTAG2-2 cells per field view that migrated through uncoated membrane (left panel) or invaded through Matrigel-coated membrane (right panel) after 24 hours. A minimum of 16 random fields were counted for each condition. F. Western blot for STAG2 protein expression in UM-UC-3 cells transduced with pEGFP (control vector) or pEGFP-STAG2 (STAG2 overexpression vector). GAPDH used as a loading control. Figure is representative of three independent experiments. G. UM-UC-3 empty vector (EV) or pEGFP-STAG2 (STAG2-overexpressed) cells seeded in the top of a Transwell insert and allowed to move through Matrigel-coated membrane (top panels, invasion) or uncoated membrane (bottom panels, migration) for 24 hours. H. Average number of UM-UC-3 EV or pEGFP-STAG2 cells per field view that migrated through uncoated membrane (left panel) or invaded through Matrigel-coated membrane (right panel) after 24 hours. I. UM-UC-3 control (pCDH) or FLAG-STAG2 cells seeded in the top of a Transwell insert and allowed to move through Matrigel-coated membrane (top panels, invasion) or uncoated membrane (bottom panels, migration) for 24 hours. J. Average number of UM-UC-3 control or FLAG-STAG2 cells per field view that migrated through uncoated membrane (left panel) or invaded through Matrigel-coated membrane (right panel) after 24 hours. A minimum of 12 random fields were counted for each experiment. Data represented as mean + SEM. Individual comparisons made using unpaired Student’s t test. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.
Article Snippet: Cells were transfected with 50nM of STAG2 siRNA (SMARTpool siGENOME siRNA, Thermo Scientific) or control siRNA (On TargetPlus nonTargeting pool, Thermo Scientific) using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA).
Techniques: Western Blot, Expressing, Transfection, Control, Membrane, Migration, Transduction, Plasmid Preparation, Over Expression
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