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Figure 5 | BMC Molecular Biology

Figure 5

From: A noncoding RNA gene on chromosome 10p15.3 may function upstream of hTERT

Figure 5

FACS analysis and phenotypes in transfectants by RNAi methods. (a) FACS analysis showing induction of more than half of the transfected populations into the preG1 phase of the cell cycle. As shown in the upper two parts, the increase in apoptotic cells in transfectants by RGM249 is shown as red circled areas. DNA content analysis revealed in the lower parts, that early cells showed typical cell proliferation profile, cells later in production showed reduced cell proliferation, and later cells also showed early signs of cell death (sub-G1), in both transfectants. In the bottom parts, the histogram with gating and the threshold were set properly for correctly stained cells in a clean machine. The large pink peak represents the green light signal from cells. The line segment flanking the peak is the gating band for cells. (b) Changes in phenotypes of HLF cells receiving LacZ shRNA or RGM249 shRNA are shown. The upper, middle, and bottom part shows the phenotype of transfectants, the result of β-Gal staining as an indicator of senescence, and EGFP expression as evidence of successful transfection, respectively. At 2–3 weeks after transfection, the transfectants were induced to undergo apoptosis (upper and bottom parts) through senescence (middle part) in the presence of both a selectable marker (neo) and RGM249 (data not shown).

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