BCL7B functions as a tumor suppressor via multiple pathways.

<p><b>A–D</b>: Nomarski (<b>A, C</b>) and DAPI (<b>B, D</b>) images of KATOIII cells transfected with control siRNA (<b>A, B</b>) and <i>BCL7B</i>-specific siRNA (<b>C, D</b>). <b>E–H</b>: Fluorescence images of KATOIII cells stained with WGA-Alexa Fluor 488 (<b>E, G</b>) and DAPI (<b>F, H</b>). These cells were transfected with control siRNA (<b>E, F</b>) or <i>BCL7B</i>-specific siRNA (<b>G, H</b>). The arrowheads indicate the presence of three nuclei in one cell. <b>I</b>: The percentages of the surviving KATOIII cells after transfection with control siRNA or <i>BCL7B</i>-specific siRNA and addition of actinomycin D. The number of living cells was counted 5 h after the addition of actinomycin D using trypan blue dye exclusion; this number was then divided by the total cell number. <b>J</b>: Percentages of KATOIII cells transfected with control siRNA or <i>BCL7B</i>-specific siRNA in the G₀/G₁, S, and G₂ phases. More than twenty thousand cells were counted for each analysis. <b>K, L</b>: The expression levels of ß-catenin (<b>K</b>) and c-FLIP (<b>L</b>) by the qRT-PCR analysis. <b>M</b>: Survival rates of KATOIII cells transfected with <i>EGFP</i>, as a control, or <i>BCL7B_EGFP</i>. The number of living cells was counted using trypan blue dye exclusion, and this number was then divided by the total cell number. <b>N</b>: The rates of apoptotic KATOIII cells transfected with <i>EGFP</i>, as a control, or <i>BCL7B_EGFP</i>. Apoptotic cells were stained for Annexin-V and/or 7-AAD and monitored by flow cytometric analysis. More than ten thousand cells were counted. Each experiment was performed three times independently. Error bars indicate SEM. The asterisks indicate the statistical significance of differences between the two groups. *p<0.05, **p<0.005, ***p<0.001. N.S.: no significance. Scale bar = 25 µm.