Figure 1.
Figure 1. Silencing of CCR2 expression by pSUPER-H1.shCCR2. (A) Expression of murine CCR2 in HEK 293 cells after cotransfection with pEGFPN-1/CCR2 and pSUPER-H1.Empty (1.5 μg), relative to that of HPRT, was not affected by pSUPER-H1.shLuc or pSUPER-H1.shInert, whereas CCR2 mRNA levels were up to 94% reduced after cotransfection with increasing amounts of pSUPER-H1.shCCR2 (***P < .001). Error bars represent SEM. (B) HEK 293 cells cotransfected with either pEGFPN-1/CCR2 and pSUPER-H1.Empty (1.5 μg; left panel) or pEGFPN-1/CCR2 and pSUPER-H1.shCCR2 (0.5 μg or 1.5 μg; middle and right panels, respectively), which show increased quenching of GFP protein expression on the cell surface, indicating almost complete loss of CCR2 surface protein after transfection with pSUPER-H1.shCCR2. Cells were viewed with a Biorad 2-photon confocal laser scanning microscope (1000× magnification), and images were digitized with Adobe Photoshop 7.0.

Silencing of CCR2 expression by pSUPER-H1.shCCR2. (A) Expression of murine CCR2 in HEK 293 cells after cotransfection with pEGFPN-1/CCR2 and pSUPER-H1.Empty (1.5 μg), relative to that of HPRT, was not affected by pSUPER-H1.shLuc or pSUPER-H1.shInert, whereas CCR2 mRNA levels were up to 94% reduced after cotransfection with increasing amounts of pSUPER-H1.shCCR2 (***P < .001). Error bars represent SEM. (B) HEK 293 cells cotransfected with either pEGFPN-1/CCR2 and pSUPER-H1.Empty (1.5 μg; left panel) or pEGFPN-1/CCR2 and pSUPER-H1.shCCR2 (0.5 μg or 1.5 μg; middle and right panels, respectively), which show increased quenching of GFP protein expression on the cell surface, indicating almost complete loss of CCR2 surface protein after transfection with pSUPER-H1.shCCR2. Cells were viewed with a Biorad 2-photon confocal laser scanning microscope (1000× magnification), and images were digitized with Adobe Photoshop 7.0.

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