Renal cystadenoma histology in the treated mice.

<p>Representative tumor images are shown for each treatment cohort, selected from the kidneys with the largest tumor volume for each. Three cystadenoma each are shown at 100×, with a portion of the tumor indicated by the frame shown at 400× (below). Cystadenomas from control mice are shown in the first column, two from IP vehicle treated mice, and one from a sucrose treated mouse. Note the cystic nature of the tumors from the rapamycin-treated mice, as well as the flattened, thin nature of the cyst-lining cells, in contrast to cystadenomas from all other mice. Arrows point to prominent enlarged but viable cells in the bortezomib-treated mice.</p

Immunoblot analysis of kidney lysates to examine therapeutic specificity.

<p><i>Tsc2⁺⁻</i> A/J strain mice were treated with vehicle, rapamycin, bortezomib or metformin for 1 week, and kidney lysates were prepared for immunoblot analysis. A) Blot strips were incubated with the antibodies against pACC-S79, AKT, pS6-S235/236, pS6-S240/244 and S6. B) Blot strips incubated with antibodies against pRaptor-S792, GRP78 and AKT.</p

Macroscopic and microscopic kidney tumor scores for <i>Tsc2^+/−</i> mice treated with five different treatment regimens.

<p>Scores are shown for each kidney available from each mouse in these cohorts. A. Macroscopic kidney tumor scores. B–C. Microscopic kidney tumor volume is shown twice, on a linear y axis scale (B) and on a logarithmic y axis scale (C). For B and C, the number of mice and kidneys examined were: 5 and 7, 9 and 15, 7 and 12, 8 and 14, and 10 and 18, respectively, for IP vehicle, rapamycin, bortezomib, sucrose, and metformin cohorts, respectively.</p

Immunohistochemistry (IHC) analysis of therapeutic effects.

<p><i>Tsc2^+/−</i> A/J strain mice of ages 9–10 months were treated with drugs for 1 week, and kidneys prepared for histology. Sections were prepared and stained using pS6-S235/236 (red), pACC-S79 (red), pIκBα-S32/34 (red) and GRP78 (red) antibodies. The bottom panel shows apoptosis assessed by TUNEL method. Representative sections are shown. All images shown are taken at 100× magnification. Insets show portions of the tumor at higher magnification (400×).</p

Body weight of mice on treatment.

<p>Body weight of mice on treatment.</p