Figure 1.

Repeated delivery of lipoplexes containing the ITSN-1–specific siRNA and myc-EH_ITSN plasmid to murine lungs produces efficient long-term ITSN down-regulation and EH_ITSN protein expression; genotyping of K0^ITSN mice. A: Lung lysates (70 μg per lane) of KD^ITSN transduced with the myc-EH_ITSN lipoplexes (days 2, 6, 12, and 18 of treatment) were subjected to Western blotting (WB) using ITSN antibody (Ab) and myc Ab. Actin was used as loading control. Wild-type (wt) mice (lane a), mice injected with empty liposomes (lane b, top panel) or empty vector (lane b, bottom panel) were used as controls. B: Conventional RT-PCR on lung samples from wt and K0^ITSN+/− mice was used to analyze ITSN-1s mRNA levels, relative to internal control, cyclophilin (cyclo, left panels). ITSN protein levels were analyzed in lung lysates (70 μg total protein per lane) of wt and K0^ITSN+/− mice by WB using ITSN Ab. Actin was used as loading control (right panels). All mice used in this study were genotyped. C and D: Densitometric analyses of ITSNand myc-EH_ITSN expression, respectively, in control mice and KD^ITSN mice transduced with the myc-EH_ITSN. Data are shown as ITSN/actin (C) or myc-EH_ITSN/actin (D) ratio. Data are expressed as means ± SEM (B–D). n = 3 (A, C, and D, mice per group); n = 3 (A, B–D, different experiments). ^∗∗P < 0.01 versus wt mice.