Fig. 4

BCL3ANT inhibits melanoma cell proliferation and migration. A Mel Ju cells were treated with DMSO or 50 μM BCL3ANT for 24 h before being stained with antibodies against Bcl-3 (red) and 406-diamidino-2-phenyl indole DAPI (blue) and imaged using confocal microscopy. B Mel Ju cells were treated with DMSO or 50 μM BCL3ANT for 24 h and the percentage of fragmented DNA and (C) cell viability were analyzed using a NucleoCounter. D Mel Ju and SK-MEL3 cells were treated with DMSO or 50 μM BCL3ANT for 24–96 h before being subjected to a WST-1 assay to measure cell proliferation. E Mel Ju cells were treated with DMSO or 50 μM BCL3ANT for 48 h and the cell cycle was assessed using a NucleoCounter. Representative histograms showing the cell cycle phases from Mel Ju cells after treatment with DMSO or BCL3ANT. F Mel Ju cells were seeded onto a 6-well plate and incubated for 24 h to adhere to the plate. A wound was created using a 1 ml pipet tip before the cells were treated for 23 h with DMSO or 50 μM BCL3ANT. An image of the wound was taken at 0 h and 23 h post treatment, and the area of the wound was calculated to determine cell migration. G NUDE mice were transplanted with SK-MEL3 melanoma cells and 6 days post-transplantation, mice were treated with 10 mg/kg BCL3ANT or vehicle control every 3 days until Day 16 when they were sacrificed. Tumor volume was measured using a CALIPER. H Six mice per group were treated with 0.04, 0.08, or 0.16 mg/kg BCL3ANT once and observed for 30 days for the development of adverse effects. All data are presented as the mean ± SEM from three independent experiments. Statistical significance was assessed using Student’s t test