Ming Hong Shen, Paulina Samsel, Louise L

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Assessment of Response of Kidney Tumors to Rapamycin and Atorvastatin in Tsc1+/− Mice Ming Hong Shen, Paulina Samsel, Louise L. Shen, Kalin Narov, Jian Yang, Julian R. Sampson Translational Oncology Volume 10, Issue 5, Pages 793-799 (October 2017) DOI: 10.1016/j.tranon.2017.07.009 Copyright © 2017 The Authors Terms and Conditions

Figure 1 MRI assessment of treatment efficacy on renal tumors of Tsc1+/− mice. Tsc1+/− mice were subjected to MRI scanning before and after treatment, respectively, in vivo. Total number and volume of all renal lesions (cystic/papillary/solid) per mouse were obtained using Analyze 10.0. Left panel: Comparison of total number and volume of renal tumors between treatment groups before starting treatment at the age of 12 months (the first MRI scan). Middle panel: Comparison of total number and volume of renal tumors between treatment groups after treatment at the age of 14 months (the second MRI scan). Tsc1+/− mice were treated with vehicle, atorvastatin (100 mg/kg), rapamycin (5 mg/kg), or combination of rapamycin (5 mg/kg) and atorvastatin (100 mg/kg) for 2 months before the second MRI scanning. Right panel: Comparison of fold change of total renal tumor number and volume between treatment groups (the second/first scan). See Supplementary Tables 1-3 for statistical details. Translational Oncology 2017 10, 793-799DOI: (10.1016/j.tranon.2017.07.009) Copyright © 2017 The Authors Terms and Conditions

Figure 2 Changes in renal tumor size of Tsc1+/− mice detected by MRI and histology. Tsc1+/− mice were scanned using MRI in vivo as in Figure 1 and sacrificed after the second scan for kidney section preparation. Kidney sections were HE-stained. Representative MRI images and HE sections were presented to show size of different tumor types at specified treatment groups. Translational Oncology 2017 10, 793-799DOI: (10.1016/j.tranon.2017.07.009) Copyright © 2017 The Authors Terms and Conditions

Figure 3 Treatment effect on proliferation of renal tumor cells in Tsc1+/− mice. Kidney sections, prepared from 14-month-old Tsc1+/− mice after the second MRI scan, were stained with antibody against Ki67 to assess proliferation of tumor cells. Representative sections were presented to show expression of Ki67 in tumor cells of specified treatment groups. Translational Oncology 2017 10, 793-799DOI: (10.1016/j.tranon.2017.07.009) Copyright © 2017 The Authors Terms and Conditions

Figure 4 Treatment effect on mTOR signaling and expression of p21 in the liver of Tsc1+/− mice. (A) Western analysis. Proteins were prepared from liver tissues dissected from Tsc1+/− mice treated for 2 months with vehicle, atorvastatin, rapamycin, or atorvastatin/rapamycin combination. β-Actin was used as a loading control. Representative Western blots were presented to show phosphorylation of Akt at S473 and S6 at S235/236 and expression of p21. (B) q-PCR analysis. Total RNA was isolated from liver tissues dissected from Tsc1+/− mice treated for 2 months with vehicle, atorvastatin, rapamycin, or atorvastatin/rapamycin combination (n=5 each group). RNA was used to synthesize cDNA, and real time q-PCR was performed to estimate relative quantity of Cdkn1a transcripts. Translational Oncology 2017 10, 793-799DOI: (10.1016/j.tranon.2017.07.009) Copyright © 2017 The Authors Terms and Conditions

Figure 5 Treatment effect on mTOR signaling of renal tumors in Tsc1+/− mice. Kidney sections, prepared from 14-month-old Tsc1+/− mice after the second MRI scan, were used for IHC analysis. Representative IHC-stained sections were presented to show phosphorylation of S6 at S235/236 and Akt at S473 in renal tumors. Black lines are scale bars. Translational Oncology 2017 10, 793-799DOI: (10.1016/j.tranon.2017.07.009) Copyright © 2017 The Authors Terms and Conditions

Figure 6 Treatment effect on MAPK signaling of renal tumors in Tsc1+/− mice. Kidney sections, prepared from 14-month-old Tsc1+/− mice after the second MRI scan, were used for IHC analysis. Representative IHC-stained sections were presented to show phosphorylation of RAF1 at S259 and Erk1/2 at T202/Y204 in renal tumors. Black lines are scale bars. Translational Oncology 2017 10, 793-799DOI: (10.1016/j.tranon.2017.07.009) Copyright © 2017 The Authors Terms and Conditions