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. 2021 Sep 25;13(19):4801. doi: 10.3390/cancers13194801

Figure 2.

Figure 2

Molecular analyses of therapy testing in the Vhl/Trp53/Rb1 mutant mouse ccRCC model. (A) Representative immunohistochemical stainings for CA9, GLUT-1, P-4E-BP1, and Ki-67 in regions of cortex and ccRCC from untreated mice and from regions of ccRCC from mice treated with PT2399, ACF, or FTY720. Scale bar = 100 mm. (B–D) Scoring of staining intensities of CA9 (B), GLUT-1 (C), and phospho-Thr37/Thr46-4E-BP1 (P-4E-BP1) (D) in ccRCC tumors. Analyses are based on 58, 25, 14, and 19 tumors for CA9, on 28, 17, 12, and 17 tumors for GLUT-1, and on 26, 24, 22, and 22 tumors for P-4E-BP1 in untreated, PT2399, ACF, and FTY720 treated cohorts. p-values were calculated using the two-sided Mann–Whitney U test without adjustments for multiple comparisons. Representative examples of scores 1, 2, and 3 for each staining are shown in Supplemental Figure S2. (E) Quantification of percentage of Ki-67 positive nuclei in regions of normal cortex (N) and ccRCC (T). Analyses are based on 22, 24, 18, and 25 tumors in untreated, PT2399, ACF, and FTY720 treated cohorts. Mean ± SEM are shown, p-values for pairwise comparisons were calculated by Student’s t-test followed by two-sided Mann–Whitney U test without adjustments for multiple comparisons.