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. 2018 Nov;188(11):2508–2528. doi: 10.1016/j.ajpath.2018.07.025

Figure 1.

Figure 1

Generation of apolipoprotein (APO) L1 transgenic mice. A: A doxycycline-inducible APOL1 (G1) construct was made by using plasmid pENTR3C as the backbone. B: The cDNA of TetOn3G and its promoter PTRE3G were used from Tet-On 3G Tetracycline-Inducible Expression Systems. The working flowchart is shown. C: To examine the time course effect of APOL1 protein expression, 4-week–old APOL1 (G1) transgenic mice were fed doxycycline (2.5 mg/kg body weight daily for 3 consecutive days and then every other day) for different periods (0, 3, 9, and 18 days). Kidneys were harvested. Proteins and RNAs were extracted. Protein blots of renal tissues were probed for APOL1 and reprobed for glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Representative gels are displayed. cDNAs were amplified for APOL1 mRNA. Renal tissues displayed the expression of APOL1 protein and mRNA in a time course manner. D: To assess the dose-response effect, 4-week–old APOL1 (G1) transgenic mice were administered different doses of doxycycline for 3 consecutive days. Kidneys were harvested, and proteins and RNAs were extracted. Protein blots of renal tissues were probed for APOL1 and reprobed for GAPDH. Representative gels are displayed. cDNAs were amplified for APOL1 mRNA. Doxycycline enhances the expression of the APOL1 protein and mRNA in a dose-dependent manner. n = 3. ∗∗P < 0.01 versus 0 and 18 days; †††P < 0.001 versus 0 day; ‡‡P < 0.01 versus 0 and 0.5 mg and §§§P <0.001 versus 0, 0.5, and 2 mg. PA, polyadenylation.