Cellular adhesion is reduced upon c-Jun knockdown. (A) Efficiency of c-Jun and JunB knockdown in ABC-DLBCL cell lines using specific shRNAs. Cell lysates were prepared from the indicated cell lines and subjected to SDS-PAGE followed by western blotting using the indicated antibodies. The densitometric analysis was performed by using ImageJ software. (B) Proliferation assay. Cells were cultured in 96-well plates and 3-(4,5-dimethylthiazol-2-yl)-2,5-dimethyltetrazolium bromide (MTT) was added to the cells 4 hours before lysis in isopropanol with 1% hydrochloric acid. The plate was read with a microplate spectrometer using a 570-nm filter. Results are presented as the mean + standard deviation (SD) of triplicate cultures. Statistical significance was evaluated by the Student t test. (C) Loss of cellular aggregation and adhesion upon Jun knockdown. OCI-Ly10 cells were cultured in Iscove modified Dulbecco medium (IMDM) medium with different concentrations of human plasma for 24 hours. The images were taken with the Olympus IX70 inverted fluorescence microscope (×20). (D) The cells attached to the bottom of the culture dish (16 hours) were quantified by using colorimetric detection. Results are shown as the mean + SD of triplicate cultures. (E) Adhesion of lymphoma cell lines to the indicated ECM proteins (4 hours) was determined by using the ECM Cell Adhesion Array Kit (Millipore). Results are shown as the mean ± SD of triplicate cultures. (F) Cell treatment with peptides containing the RGD or RGE sequence (30 minutes) was followed with the cell adhesion assay. OD, optical density; FNK, fibronectin; RGD, Arg-Gly-Asp motif.