2.4. Xenograft Experiments

Lung cancer A549 (3 × 10⁶) cells were subcutaneously (s.c.) inoculated into the hind leg flanks of 7-week-old Nude-Foxn1nu Hsd: athymic male nude mice (Envigo, Jerusalem, Israel). When the tumor volume reached 60–70 mm³, the mice were divided into two groups: one group was intratumorally treated with si-hVDAC1-2A mixed with the in vivo JetPEI reagent to a final concentration of 200 nM (calculated according to the tumor volume, 2 boluses) every 3 days. When the mice were sacrificed, the tumors were excised and processed for immunohistochemistry, or frozen in liquid nitrogen for immunoblotting and RNA isolation, as described in Text S1. Experimental protocols used were approved by the Institutional Animal Care and Use Committee.