Perturbing the Normal Level of SIDT1 Suppresses the Naked ASO Effect

Figure 2

The effect of SIDT1 on total cellular uptake of naked A-15. (a) MEFs were transfected with the empty plasmid (control) or pCI-neo-SIDT1 for 48 hours, and the SIDT1 expression level was analyzed by Western blotting. (b) MEFs were transfected with the empty plasmid (control) or pCI-neo-SIDT1. After the 48-hour culture, the cells were rinsed with PBS and further cultured in the absence or presence of 500 nM of naked A-15 for 24 hours. After washing the cells with PBS, they were analyzed by flow cytometry. In the left and middle panels, yellow and red histograms are from MEFs cultured in the absence and presence, respectively, of A-15. In the right panel, the red histograms in the left and middle panels are overlaid and shown in yellow and red, respectively. (c) MEFs were transfected with the control siRNA (siControl) or the siRNA against the SIDT1 mRNA (siSIDT1#1 or siSIDT1#2) for 72 hours, and the SIDT1 expression level was analyzed by Western blotting. (d) MEFs were transfected with siControl, siSIDT1#1, or siSIDT1#2. After the 72-hour culture, the cells were rinsed with PBS and further cultured in the absence or presence of 500 nM of naked A-15 for 6 hours. After washing the cells with PBS, they were analyzed by flow cytometry. In the upper panels, yellow and red histograms are from MEFs cultured in the absence and presence, respectively, of A-15. In the lower left panel, the red histograms in the upper left and middle panels are overlaid and shown in yellow and red, respectively. In the lower right panel, the red histograms in the upper left and right panels are overlaid and shown in yellow and red, respectively.

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