Expression of wild type and mutant HA-tagged ABCA1 variants in HEK293H and MDCKII cells. A. The mRNA expression level of ABCA1 was measured by real time RT-PCR in the parental HEK293H and MDCKII cells, as well as in cells expressing the ABCA1 variants. The ABCA1 expression of HepG2 cells were used as a reference for relative mRNA expression levels. For comparison, ABCA1 mRNA level was also determined in PMA-pretreated Thp-1 cells and in human monocyte-derived macrophages (MDM). To induce ABCA1 expression, the latter two cell types were treated with 1 μM T0901317, an LXR agonist (LXR). Control: original, parental, or non-treated cells; n.d. - not detected. The expression of ABCA1 variants were similar in the established cell lines and comparable with that seen in LXR-induced macrophages. B. The protein expression level of HA-tagged ABCA1 variants was determined using anti-HA antibody by Western blot analysis on whole cell lysates. Panels demonstrate representative blots for the variants of HEK293H and MDCKII cells at passage number 5. Each lane contains 15 μg and 40 μg of protein per well from HEK293H and MDCKII cells, respectively. Anti-Na/K ATPase antibody was used to control the sample loading. For further details see legend for Figure 1.
Kasza et al. BMC Cell Biology 2009 10:93 doi:10.1186/1471-2121-10-93