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. 2005 Sep 15;34(1):28–38. doi: 10.1165/rcmb.2005-0172OC

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Figure 4. — Generation of HIMF overexpressing mouse lung epithelial cells. Confluent monolayers of MLE-12 cells were transfected by HIMF cDNA or control vector with Lipofectamine 2000. Stable cell lines, MLE-HIMF and MLE-Zeo, were screened based on resistance to Zeocin (400 μg/ml). (A) Western blot from culture medium indicated that MLE-HIMF cells produce higher level of HIMF protein than their parent and transfection counterparts. (B) RT-PCR demonstrated that MLE-HIMF cells have overexpressed HIMF mRNA, and enhanced SP-B and SP-C mRNA levels compared with their parent and transfection controls. (C) Luciferase assay indicated that the promoter activity of SP-B and SP-C in MLE-HIMF cells was enhanced compared with that of their parent and transfection controls (all groups are in triplicate). * Significant increase from MLE-12 parent; ^# significant increase from transfection controls (P < 0.05). Shown is one representative experiment of three, all with similar results.

Figure 4. — Generation of HIMF overexpressing mouse lung epithelial cells. Confluent monolayers of MLE-12 cells were transfected by HIMF cDNA or control vector with Lipofectamine 2000. Stable cell lines, MLE-HIMF and MLE-Zeo, were screened based on resistance to Zeocin (400 μg/ml). (A) Western blot from culture medium indicated that MLE-HIMF cells produce higher level of HIMF protein than their parent and transfection counterparts. (B) RT-PCR demonstrated that MLE-HIMF cells have overexpressed HIMF mRNA, and enhanced SP-B and SP-C mRNA levels compared with their parent and transfection controls. (C) Luciferase assay indicated that the promoter activity of SP-B and SP-C in MLE-HIMF cells was enhanced compared with that of their parent and transfection controls (all groups are in triplicate). * Significant increase from MLE-12 parent; ^# significant increase from transfection controls (P < 0.05). Shown is one representative experiment of three, all with similar results.