Figure 2
MYCNkd induces CCL2 expression in neuroblastoma cells and NKT cell chemoattraction.
SK-N-BE(2) and LA-N-1 MYCN-amplified neuroblastoma cell lines were stably transduced with MYCN shRNA using a U6 expression cassette into pHIV-7-EGFP lentiviral plasmid. (A) Representative Western blot analysis of MYCN protein in the nuclear extracts from indicated cells. Lane 1, parental; lane 2, vector control; lane 3, MYCN shRNA. (B) CCL2 RNA expression was quantified by TaqMan RT-PCR; values are relative to expression in parental cells. Results are mean ± SD from 3 experiments. (C) CBA analysis was performed with supernatants of vector control or MYCN shRNA-transduced SK-N-BE(2) cells as described in Figure 1C. (D) Migration of freshly isolated PBLs to the neuroblastoma cell supernatants with the indicated conditions was performed as described in Figure 1C. Where indicated, supernatants were pretreated for 1 hour with neutralizing anti-CCL2, anti-CXCL8, or isotype control (IgG1) mAb. Results are mean ± SD from 3 experiments.
