Fig. 7
From: ABCB1 protects bat cells from DNA damage induced by genotoxic compounds
ABCB1 expression and efflux capability in human and multiple bat species. a qRT-PCR for ABCB1 mRNA expression in cell lines derived from human and P. alecto. ABCB1 mRNA expression is normalised to GAPDH and presented relative to the expression in WI-38 cells (mean ± SDs of three independent experiments). b Western blot analysis of ABCB1 in the indicated cell lines. Tubulin was used as a loading control. c qRT-PCR for ABCB1 mRNA expression in tissues derived from human (N = 1) and P. alecto (N = 3). ABCB1 mRNA expression is normalised to GAPDH and presented relative to the respective human tissues (mean ± SDs). d Western blot analysis of ABCB1 protein in the indicated tissues. Ponceau S staining of the membrane is shown as a loading control. Arrow indicates the size of ABCB1 protein. H and P. a indicate human and P. alecto respectively. Lysates from HEK293T transfected with control (−) or ABCB1 expressing plasmid (+) serve as negative and positive control, respectively. e Western blot analysis of ABCB1 in cells derived from the different bat species. Actin was used as a loading control. f Flow cytometry analysis of doxorubicin accumulation in cells derived from different bat species. Cells were treated and analysed as in Fig. 3b. g Western blot analyses of γH2AX in cells derived from different bat species. Cells were treated as in Fig. 4. Blots are representative of three independent experiments. h Flow cytometry analysis of doxorubicin accumulation. Cells were pre-treated without or with 5 μM verapamil (Vera) for 30 min before the treatment with 10 μM doxorubicin alone or together with Vera for an additional 3 h. The fold change by Vera treatment is presented as the mean fluorescence intensity of cells treated with both verapamil and doxorubicin relative to the mean fluorescence intensity of cells treated with doxorubicin alone. Bars represent mean fold change ± SD of at least three experiments. Data shown in Figs. 3b, c and 7f are also included in this histogram. Statistical significances were calculated using unpaired student’s two-sided t test. **p < 0.01, ***p < 0.001
