Figure 3.

Functional uptake in MHT Cells is saturable. (A) MHT cells were incubated with increasing concentrations of SR-B1 SSO alone or the indicated concentration of SR-B1 SSO and increasing concentration of control (competitor) SSO. Twenty-four hours after adding the SSO to the cells, reduction in SR-B1 mRNA was measured by qRT–PCR. (B) DNA containing competitor SSOs are effecting functional uptake of SR-B1 SSO. MHT cells were incubated with 1 µM SR-B1 SSO in the presence of 10 µM competitor SSOs with increasing gap length. The more DNA present in the gap, the better the competition. (C) MHT cells and primary hepatocytes (D) were incubated with 100 nM SR-B1 SSO alone or in the presence of 5 µM competitor SSO of different lengths. The competitor SSOs had 2 MOE nucleotides on the 5′- and 3′-ends to help protect against nuclease degradation. The length of the DNA in the gap was varied as indicated. Twenty-four hours after adding the SSOs to the cells, SR-B1 mRNA was measured with qRT–PCR. SSOs with more DNA content were found to be better competitors. (E) Effect of different length competitors on functional uptake of SR-B1 SSO. MHT cells were incubated with 1 µM of SR-B1 SSO in the presence of 1 µM competitor SSOs. The length of the SSOs was varied by increasing the MOE nucleotides on the 5′- and 3′ wings with the number of DNA nucleotides in the center gap held constant at 10 nt. Mean values and SD were measured from triplicate samples, *P < 0.01, **P < 0.05, unpaired Student’s t-test.