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A2058 tumor spheroids were seeded in 96-well plates coated with 1% agarose in PBS and cultured on a shaker at 90 rpm for 48 h. After spheroid formation, cells were infected in the following groups: mock, Ad-SV (100 MOI), or Ad-MMP-2 (25, 50, and 100 MOI). After 24 h, spheroids were transferred to 8-well chamber slides, and allowed to grow for 72 h. Cold methanol fixation was performed and followed by HEMA staining.
