GATA3 expression in classical Hodgkin lymphoma cell lines is effectively altered by small hairpin RNA (shRNA) knockdown. (A) Immunoblots of Hodgkin lymphoma cell lines L-428, L-591, L-1236, KM-H2, HDLM-2, U-HO1, and DEV. Same amount of protein (100 μg) were assayed for protein expression of GATA3. Jurkat T cells were used as positive controls. Actin was used as loading control to demonstrate equal loading. (B) Protein lysates of cell lines treated with lentiviral particles for 2 functional shRNAs (siGATA3 no. 1 and no. 2, lanes 3 and 4) and 1 control scrambled shRNA (siSCR, lane 2) for GATA3 were immunoblotted for GATA3 and actin expression. Lysates were prepared 3 days (L-1236), 4 days (U-HO1), or 5 days after infection (L-428, KM-H2). (C) Three days after infection, GFP⁺ cells were sorted. After RNA and complimentary DNA preparation, semiquantitative PCRs for GATA3 and GAPDH mRNAs were performed. In the No RT control, RNA of untreated cells was deposited into the complimentary DNA reaction without RT enzyme. A representative experiment of at least 3 independent repeated experiments is shown.