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Figure 6 | BMC Molecular Biology

Figure 6

From: Gene transfer of Hodgkin cell lines via multivalent anti-CD30 scFv displaying bacteriophage

Figure 6

Specific gene delivery of CD30 positive cell lines. Gene delivery specificity was demonstrated via competition experiment. (a) L1236 cells were incubated with M13-pAK8-Ki-4(EGFP) phagemid particles in the absence or presence of monoclonal antibody (moab) Ki-4. Representative sections were photographed with 10 × objective using fluorescence microscopy. Transduction efficiency was quantified by FACS analysis after 96 hours. Indicated cell lines were incubated with M13-pAK8-Ki4(EGFP), M13-pCANTAB-Ki4(EGFP) or M13-pAK8(EGFP) phagemid particles, respectively. (b) A homogenous cell population (P1) was gated in forward (FSC-H) by right-angle light (SSC-H) scatter plots and subsequently gated for living cells (P2) in FSC-H by propidium iodide (PI-A) channel dot plot. The living cells were finally analyzed for EGFP-expression in PI-A versus green fluorescence (FITC-A) channel dot plot, setting the gate at 0.05% of transduced cells incubated with the control phagemid particles M13-pAK8(EGFP). (c) Representative analysis of the L1236 cell line, incubated with M13-pAK8(EGFP), M13-pCANTAB6-Ki4(EGFP) or M13-pAK8-Ki4(EGFP) phagemid particles, respectively. Cells were analyzed for EGFP expression as described above. (d) Summarized results of transduction experiments with different cell lines in percentage of EGFP-expressing cells. Statistical significance was analysed according to the Student's one-tailed t test. p-values < 0.05, < 0.01 and < 0.001 were indicated with *, ** or ***, respectively.

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