Figure 1. Comparison of SLC41A3 protein levels in various HEK293 cell lines.

HEK293 WT cells (HEK WT) or the HEK293 cell line with stably integrated Tet-repressor (Cl2) were transiently transfected with pcDNA5TO-SLC41A3 (− and +) or the empty vector control (E). Protein expression in the transiently transfected cell lines (HEK WT and Cl2) and in the stably transfected cell line (A3st) was induced by addition of tetracycline to a final concentration of 1 μg/ml (+). Uninduced control cells remained untreated (−). Cells were harvested after 24 hours of induction and lysed in RIPA buffer. Total protein extracts were analyzed on an 8.5% PAA-SDS gel and immunodetection of SLC41A3 (55 kDa) was either performed with an antibody directed against the Strep-tag (a) or against the native protein (b). The signals for the monomeric forms of native and Strep-tagged SLC41A3 are indicated. The ribosomal protein RPL19 (23 kDa) served as loading control and was detected by reprobing the membranes with the respective antibody.