EMBO/soda-nel huggingface dataset:
Small molecules enhance autophagy and reduce toxicity in Huntington's disease models (2007)
figf4

(a–c) Clearance of A53T α-synuclein in stable PC12 cells as in Figure 1b—treated with DMSO (control), or with 0.2 μM rapamycin alone, SMER alone (140 μM SMER10 (a), 43 μM SMER18 (b) or 47 μM SMER28 (c)) or both for the 8 h switch-off period—was analyzed by immunoblotting with antibody against HA (above) and densitometry analysis relative to actin (below). The concentration of rapamycin is saturating for its effect on the clearance of A53T α-synuclein. Error bars denote s.e.m. P = 0.0025 (Rap), P = 0.0018 (SMER10), P 0.0001 (SMER10 + Rap), P 0.0001 (Rap or SMER10 versus SMER10 + Rap) (a); P = 0.0069 (Rap), P = 0.0498 (SMER18), P 0.0001 (SMER18 + Rap), P = 0.0038 (Rap versus SMER18 + Rap), P = 0.0007 (SMER18 versus SMER18 + Rap) (b); P 0.0001 (Rap, SMER28, Rap versus SMER28 + Rap, SMER28 versus SMER28 + Rap) (c). (d–f) The percentage of EGFP-positive cells with EGFP-HDQ74 aggregates (above) and cell death (below) in COS-7 cells, as in Figure 1c—treated with DMSO (control), or with 0.2 μM rapamycin alone, SMER alone (140 μM SMER10 (d), 43 μM SMER18 (e) or 47 μM SMER28 (f)) or both for 24 h—were expressed as odds ratios. Error bars: 95% confidence interval. Aggregation (top panels): P = 0.248 (Rap), P = 0.217 (SMER10), P 0.0001 (SMER10 + Rap), P 0.001 (Rap or SMER10 versus SMER10 + Rap) (d); P = 0.248 (Rap), P = 0.543 (SMER18), P 0.0001 (SMER18 + Rap), P = 0.008 (Rap versus SMER18 + Rap), P = 0.002 (SMER18 versus SMER18 + Rap) (e); P = 0.248 (Rap), P = 0.002 (SMER28), P 0.0001 (SMER28 + Rap), P 0.0001 (Rap versus SMER28 + Rap), P = 0.012 (SMER28 versus SMER28 + Rap) (f). Cell death (bottom panels): P = 0.002 (Rap), P 0.0001 (SMER10, SMER10 + Rap, Rap or SMER10 versus SMER10 + Rap) (d); P = 0.002 (Rap), P = 0.948 (SMER18), P 0.0001 (SMER18 + Rap), P = 0.015 (Rap versus SMER18 + Rap), P 0.0001 (SMER18 versus SMER18 + Rap) (e); P = 0.002 (Rap), P 0.0001 (SMER28, SMER28 + Rap, Rap or SMER28 versus SMER28 + Rap) (f). Note that we have treated cells for a shorter time in this experiment (24 h), compared with Figure 1c (48 h). This probably accounts for the failure of the protective trends of rapamycin and some of the SMERs to reach significance for aggregation on their own in some of the experiments. ***P 0.001; **P 0.01; *P 0.05; NS, nonsignificant.
Small molecules enhance autophagy and reduce toxicity in Huntington's disease models (2007)
figf5

(a–c) Clearance of A53T α-synuclein in stable PC12 cells as in Figure 1b—treated for 24 h with either DMSO (control), or with 47 μM SMER10 and its analogs (SMER10a–c) (a), 43 μM SMER18 and its analogs (SMER18a–l) (b), or 47 μM SMER28 and its analogs (SMER28a–k) (c)—was analyzed by immunoblotting with anti-HA antibody (above) and densitometry analysis relative to actin (below). All the analogs were used in the cell culture medium at 1:400 dilution of 5 mg ml−1 stock solution (in DMSO). Error bars denote s.e.m. P = 0.0058 (SMER10a), P = 0.6736 (SMER10b), P = 0.9507 (SMER10c), P = 0.0481 (SMER10) (a); P = 0.0006 (SMER18a), P = 0.0249 (SMER18b), P = 0.0167 (SMER18c), P = 0.0117 (SMER18d), P = 0.0269 (SMER18e), P = 0.0165 (SMER18f), P = 0.0148 (SMER18g), P = 0.0011 (SMER18h), P = 0.7369 (SMER18i), P = 0.0012 (SMER18j), P = 0.1531 (SMER18k), P = 0.0006 (SMER18l), P = 0.0001 (SMER18) (b); P = 0.0014 (SMER28a), P = 0.0002 (SMER28b), P = 0.0001 (SMER28d), P = 0.0048 (SMER28h), P = 0.0002 (SMER28i), P = 0.0162 (SMER28j), P 0.0001 (SMER28c, SMER28e–g, SMER28k, SMER28) (c). (d–f) The percentage of EGFP-positive cells with EGFP-HDQ74 aggregates in COS-7 cells as in Figure 1c—treated for 48 h with either DMSO (control), or with 47 μM SMER10 and its analog (SMER10a) (d), 43 μM SMER18 and its analogs (SMER18a, SMER18c–h, SMER18j, SMER18l) (e), or 47 μM SMER28 and its analogs (SMER28a–k) (f)—were expressed as odds ratios. All the analogs were used in the cell culture medium at 1:400 dilution of 5 mg ml−1 stock solution (in DMSO). Error bars: 95% confidence interval. P = 0.003 (SMER10a), P = 0.004 (SMER10) (d); P 0.0001 (SMER18a, SMER18c, SMER18d, SMER18f, SMER18h), P = 0.009 (SMER18e), P = 0.001 (SMER18g), P = 0.382 (SMER18j), P = 0.067 (SMER18l), P = 0.031 (SMER18) (e); P 0.0001 (SMER28a, SMER28c, SMER28e–g, SMER28i–k, SMER28), P = 0.574 (SMER28b), P = 0.041 (SMER28d), P = 0.002 (SMER28h) (f). ***P 0.001; **P 0.01; *P 0.05; NS, nonsignificant.
Rebuilding core abscisic acid signaling pathways of Arabidopsis in yeast (2019)
Figure 1

Fig. 1. Reconstruction of the ABA signal pathway in yeast.

The SnRK2 OST1 activates ABFs for expression of a luciferase reporter (LUC) in yeast. Luciferase activity is given on a Log10 scale as relative counts of light emission per second (rcps) normalized to the cell density. The OST1-mediated induction factor is indicated above the columns. In the absence of ABFs, the LUC activity was 3 x 103 rcps.

Combining the ABA signaling components ABF2, OST1, the PP2C ABI1, and RCAR11 provides ABA–controlled LUC expression. The reporter is under the control of a minimal promoter that contains ABA-responsive cis elements (ABREs). Core ABA signaling involves the RCAR receptor that inhibits the ABA coreceptor (PP2C) in the presence of ABA and allows the activation of the protein kinase SnRK2. SnRK2 phosphorylates and activates the key transcription factor ABF, which targets promoters containing ABRE. Reporter activity was determined 16 h after exogenous ABA (0.1 mM) administration and referred to samples expressing ABF2 alone.

Data information: The bar-plot with single data points represents the mean ± s.e.m.; n = 6 biological replicates derived from three independent yeast transformants.

Source data are available online for this figure.
Rebuilding core abscisic acid signaling pathways of Arabidopsis in yeast (2019)
Figure 2

Fig. 2. Activation of ABA signaling by the Arabidopsis receptor family members in yeast and plant protoplasts.

In yeast, the OST1- and ABF2-mediated reporter activity was inhibited by ABI1 (‑RCAR). The subsequent recovery of the response was determined by co-expressed ABA receptors RCAR1-RCAR14 belonging to three subfamilies, in the presence or absence of 30 µM ABA. The ABA response is expressed relative to the response obtained by replacing ABI1 with the catalytically inactive ABI1D177A.

Regulation of an ABA-responsive LUC reporter in Arabidopsis protoplasts by ectopic expression of 1 µg ABI1-effector in combination with 5 µg ABA receptor DNA (± 10 µM ABA) per sample containing 105 protoplasts.

Data information: In (A) each bar represents the mean ± s.e.m; n = 12, biological replicates (derived from four independent yeast transformants); for RCAR1/RCAR5 and RCAR3/RCAR13 18 and 15 biological replicates were used, respectively. Outliers detected by Grubbs-test were removed from the analysis. In (B) each bar represents the mean ± s.e.m of three independent transformations normalized to the activity of a control samples without RCAR and ABA.

Source data are available online for this figure.
Rebuilding core abscisic acid signaling pathways of Arabidopsis in yeast (2019)
Figure 3

Fig. 3. Comparison of ABA coreceptor regulation by RCAR12 and RCAR13 in yeast, Arabidopsis protoplasts, and in vitro.

The ABA-dependent regulation of the PP2Cs ABI1 (A,D,G), ABI2 (B,E,H), and HAB1 (C,F,I) by RCAR12 and RCAR13 were analyzed in yeast (A-C), Arabidopsis protoplasts (D-F), and in vitro (G-I).

A-C) The analysis in yeast was conducted as mentioned in Fig. 2A except that other PP2C – RCAR combinations were used and the ABA concentrations varied as indicated.

D-F) Analysis of protoplasts was performed in the ABA-deficient background aba2-1 in the presence of different exogenous ABA concentrations. The ABA response was approximately tenfold inhibited by ectopic expression of the respective PP2Cs (Tischer et al, 2017). The induction of the reporter gene is related to protoplasts without ectopic RCAR expression, and in the absence of exogenous ABA.

G –I) The in vitro analysis was performed by using 50 nM PP2C and 100 nM RCAR at different ABA concentrations. Maximum inhibition of PP2C activity by ABA was set to 100%.

Data information: In (A-C) each data point represents the mean ± s.d.; n = 9 biological replicates derived from three independent yeast transformants. In (D-F) data represent the mean ± s.d. of three independent transfections. In (G-I) data represent the mean ± s.d. of three replicates.

Source data are available online for this figure.
Rebuilding core abscisic acid signaling pathways of Arabidopsis in yeast (2019)
Figure 4

Fig. 4. Regulation of the ABA response by different protein kinases of the SnRK2 family in yeast.

Dendrogram of the Arabidopsis SnRK2 protein kinases belonging to three subgroups.

Expression of the SnRK2s in yeast revealed ABF2-mediated transactivation by SnRK2.1, SnRK2.4, SnRK2.5, and SnRK2.10 in addition to OST1. Inset: Immuno-detection of FLAG-tagged SnRK2s expressed in yeast.

The active subgroup I members together with SnRK2.2 and SnRK2.3 were analyzed for differential activation of ABFs. The OST1 data shown in Fig. 1A is included for better comparison. The induction level of ABF4 relative to the empty vector control (ev) is indicated.

The OST1 domain structure comprises the kinase domain (KD) containing the activation loop and the two carboxyterminal regulatory domains DI and DII/ABA-box. Potential phosphorylation sites within the activation loop are underlined.

Increased ABF2-mediated reporter expression by OST1 harboring an ABA-box deletion as indicated in D).

Analysis of the phosphorylation status in the activation loop of SnRK2s expressed in yeast. Phosphorylation occurred at serine 175 or threonine 176, and at threonine 179. The relative abundance is given as the ratio of signal intensity of phosphorylated versus non-phosphorylated peptide for OST1, SnRK2.3, and SnRK2.4 in the mass spectrometric analysis. The numbering of phosphosites corresponds to OST1. The peptide sequences of the SnRK2 activation loop and the details for phosphosite detection and quantification are provided in Appendix Figure S3 and Fig. EV 5.

Inhibition of OST1 and SnRK2.4 by clade A PP2Cs.

Data information: The data in (B, C, E, G) represents the mean ± s.e.m; n = 6 biological replicates derived from three independent yeast transformants. The data in (B) and (E) are normalized to control samples expressing ABF2 but no SnRK2 and (G) to cells expressing inactive ABI1D177A instead of ABI1. Data in (F) represents the mean ± s.d. of four biological replicates.

Source data are available online for this figure.
Rebuilding core abscisic acid signaling pathways of Arabidopsis in yeast (2019)
Figure 5

Fig. 5. Different ABA sensitivity conferred by SnRK2 members in yeast.

A,B) ABF2-mediated reporter induction by different SnRK2s was assessed in the presence of the receptor complex RCAR11 and ABI1, and varying exogenous ABA concentrations.

C) Maximum ABA response recovery of samples shown in (A,B) relative to the reporter activity of 0.1 mM ABA-treated yeast cells expressing SnRK2s but no ABI1.

D-F) Analysis of the ABA response mediated by OST1 and SnRK2.4 using the receptors RCAR1 (D), RCAR8 (E), and RCAR14 (F).

Data information: The ABA concentration that provides half maximum response recovery is indicated by dashed vertical lines. The data in (A-C) represent the mean ± s.d.; n = 6 biological replicates derived from three independent yeast transformants. Replicates were normalized to controls without SnRK2 in (A, B) and to controls without ABI1 and RCAR for each SnRK2 in (C). The data in (D-F) are the mean ± s.e.m; n = 12, biological replicates from four independent yeast transformants for OST1 + RCAR1/RCAR8, and SnRK2.4 + RCAR8. The other combinations were analyzed in nine biological replicates derived from three independent yeast transformants.

Source data are available online for this figure.
Rebuilding core abscisic acid signaling pathways of Arabidopsis in yeast (2019)
Figure 6

Fig. 6. Comparison of OST1 and SnRK2.4 in the interaction with ABI1 affected by ABA receptors and ABA.

FRET-FLIM analysis of GFP:OST1 or GFP:SnRK2.4 and the effect of ABI1 fused to the mCherry fluorophore in the presence of RCAR1 and ABA. GFP lifetimes were determined after transient expression of the Arabidopsis proteins in leaves of Nicotiana benthamiana.

Representative false colour images of GFP:SnRK2 lifetimes measured in leaf tissues expressing OST1 or SnRK2.4 in combination with mCherry:ABI1 and RCAR1 without exogenous ABA. The bar marks 20 µm and the nucleus (n) and cytosol (c) are indicated.

Analysis of SnRK2-ABI1 interaction as mentioned in (A) but in the presence of RCAR14 and different exogenous ABA levels. RCAR14 requires ABA for efficient displacement of SnRK2s from ABI1.

Representative GFP lifetime false colour image of the analysis shown in C) 2h after 30 µM exogenous ABA administration. Details are as described in (B).

Data information: The single data points shown in (A and C) were collected from three independent experiments. Details about the analysed n and p-values are provided in the source data file. Boxes show the quartiles and the mean. Stars indicate significance of two-tailed student’s t‑test with p < 0.05 (*),< 0.001 (***).
Synaptic disruption and CREB-regulated transcription are restored by K+ channel blockers in ALS (2021)
Figure 1

Figure 1. Altered transcriptional program and CREB activation in ALS.

(A) Heatmap representing the differentially expressed genes in ALSC9orf72 (ALSC9orf72 I and ALSC9orf72 II lines) vs Healthy (Healthy I and CorrectedC9orf72 lines) MN transcriptomes. Colour scale represents gene expression relative to the Healthy genotype.

(B-C) GO (biological processes) terms of the top significant down- or up-regulated pathways in ALSC9orf72 MN (hypergeometric test).

(D) Reactome representation of clustering and distribution of the significantly up- and down-regulated pathways in ALSC9orf72 MN.

(E) PASTAA ranking of the top 3 transcription factors involved in the regulation of the differentially expressed genes in ALSC9orf72 cultures. Significance was set with the algorithm published in Roider et al. (2009).

(F) Single tube qPCR experiments showing the down-regulation of the synaptic genes NLGN3, SLC6A, TRIM9, SYT1, SYNGR1, and SNAP91 in ALSC9orf72 MN (Welch's t-test). n=3 independent cultures for each hiPSC line.

(G-H) Time course analysis of nuclear pCREBS133 levels and MN survival in ALSC9orf72 and Healthy cultures (two-way ANOVA). n=3 independent cultures for each hiPSC line (for each time point). Scale bar: 25 µm.

(I) Representative confocal image showing colocalization of the CREB interactor CBP within cytotoxic aggresomes. Scale bar: 5 µm. Dashed line represents the cell soma.

Data information: *p<0.05; **p<0.01; ***p<0.001. Error bars represent SEM. Arrow indicates the structure displayed at higher magnification. Exact p-values are reported in Appendix Table S1.
Synaptic disruption and CREB-regulated transcription are restored by K+ channel blockers in ALS (2021)
Figure 2

Figure 2. Neuroprotective effect of Apamin and XE991 in ALSC9orf72 MN.

(A) Both K+ channel blockers exert a neuroprotective effect in ALSC9orf72 cultures by increasing the neurite length of mutant MN (one-way ANOVA followed by Dunnett's multiple comparison test). n=3 independent treatments with each hiPSC line. Scale bar: 50 µm.

(B) Representative confocal image showing autophagic targeting of cytotoxic aggresomes in ALSC9orf72 MN upon K+ channel blockade. Scale bar: 5 µm. Dashed line represents the cell soma.

(C) Apamin and XE991 decrease the size of aberrant aggresomes, confirming enhanced autophagy degradation (one-way ANOVA followed by Dunnett's multiple comparison test). n=3 independent treatments with the ALSC9orf72 I line. Scale bar: 5 µm.

(D) K+ channel blockers increase the levels of autophagy-specific phospho-SQSTM1/p62S403 in ALSC9orf72 MN (one-way ANOVA followed by Dunnett's multiple comparison test). n=3 independent treatments with the ALSC9orf72 I line.

(E) The load of aggregated SQSTM1/p62 is also reduced by the treatments in ALSC9orf72 cultures (one-way ANOVA followed by Dunnett's multiple comparison test). n=3 independent treatments for each cell line. Scale bar: 20 µm.

(F) Treatment with the K+ channel blockers reduces the number of GGGGCC toxic RNA foci in ALSC9orf72 MN. n=3 independent treatments with each hiPSC line (one-way ANOVA followed by Dunnett's multiple comparison test). Scale bar: 5 µm. Dashed line represents the cell soma.

Data information: *p<0.05; **p<0.01; ***p<0.001. Error bars represent SEM. Arrow indicates the structure displayed at higher magnification. Exact p-values are reported in Appendix Table S1.
Synaptic disruption and CREB-regulated transcription are restored by K+ channel blockers in ALS (2021)
Figure 3

Figure 3. The neuroprotective effect of the K+ channel blockers includes restoration of CREB-controlled transcription and of synapse composition.

(A-B) Volcano plots obtained by DESeq2 analysis showing the effect on the transcriptome of ALSC9orf72 MN exerted by Apamin and XE991 in the ALSC9orf72 II and ALSC9orf72 III lines.

(C) PASTAA ranking of the top transcription factors activated by Apamin and XE991. Significance was set with the algorithm published in Roider et al. (2009).

(D) Venn diagram and showing the shared 263 CREB-controlled genes significantly altered by treatment with K+ channel blockers; the bar plot represents the significantly altered genes involved in neuronal activity and autophagy obtained by DESeq2 analysis.

(E) Apamin and XE991 increase the levels of pCREBS133 in all the ALSC9orf72 lines (one-way ANOVA followed by Dunnett's multiple comparison test). n=3 independent treatments with each hiPSC line. Scale bar: 10 µm.

(F) ALSC9orf72 MN showed an increased number of Homer1b/c+ synaptic contacts after treatment with the K+ channel blockers, which in contrast had no significant effect on Healthy MN. n=3 independent treatments with each hiPSC line (two-way ANOVA). Scale bar: 5 µm.

(G) Venn diagram showing the number of proteins significantly altered upon Apamin and XE991 treatment in ALSC9orf72 MN. The lines ALSC9orf72 II and ALSC9orf72 III were used as representative of the ALS genotype.

(H-I) Volcano plot displaying the up- and down-regulation of the proteins altered by Apamin and XE991 (t-test). The blue dots indicate proteins involved in synaptic structure and function.

(J) Up-regulated GO (biological processes) terms in ALSC9orf72 MN treated with either K+ channel blockers (Fisher enrichment test).

(K) Synapse- and neuronal processes-related proteins significantly up-regulated by both treatments.

Data information: *p<0.05; **p<0.01. Error bars represent SEM. Exact p-values are reported in Appendix Table S1.

Expanded View Figure Legends
Kidney Cancer PDOXs reveal patient-specific pro-malignant effects of antiangiogenics and its molecular traits (2020)
Figure 1

Figure 1. Effects of anti-VEGF/R therapy on tumor progression, invasiveness and metastatization in 786-O orthoxenograft.

When tumor was palpable, mice were treated with two different antiangiogenics, DC and Beva. For short treatment experiments, the duration of treatment was 14 days. For survival experiments, therapies were administered until survival of each mouse.

B-F) Effects of DC and Beva treatment in 786-O orthoxenograft. Evaluation of (B) tumor progression (average±SD), (C) tumor weight at sacrifice, (D) vessel number, (E) percentage of necrotic area by histology, and (F) survival by Kaplan Meier plot in control and treated animals. Box plots indicate median, Q1/Q3 and max/min value whiskers in Control (black), DC101 (red) and Beva (blue) treatment groups. 5-8 tumors/group were analyzed by Mann-Whitney test, and Mantel-Cox test for survival where *p < 0.05, **p < 0.01 and ***p < 0.001 and DC * p < 0.05 and Beva # p < 0.05 vs Control).

G) Representative invasive front profile of 786-O control and short-term DC and Beva treated tumors in vimentin stained sections (4X). Front (red line) and rear (white line) edges of tumor protrusion into kidney determine the depth of invasion (green arrow) perpendicularly connecting red and white lines. Box plot shows the quantification of invasion indicating median, Q1/Q3 and max/min value whiskers of fold-invasion (∆ invasion) in treated vs control tumors (7 samples/group; *p < 0.05 by Mann-Whitney test).

H) Evaluation of lung micrometastases in survival experiments. Incidence (pie chart, *p < 0.05 by Chi-square) and proportion of mice with 0, 1, 2 or > 3 metastases (mets) per lungs (*p < 0.05 by Mann-Whitney test) are shown.
Kidney Cancer PDOXs reveal patient-specific pro-malignant effects of antiangiogenics and its molecular traits (2020)
Figure 2

Figure 2. Establishment and histological characterization of Ren-PDOX.

Representation of protocol for the establishment of Ren-PDOXs. Ren-PDOXs were obtained by implantation of a total of 56 ccRCC human specimens on left mouse kidney. Representative images of Ren-PDOX tumor and lung metastases (mets, arrows) are shown. Total number of established and metastatic Ren-PDOXs is reported. Graph describes the proportion of metastatic Ren-PDOXs generated by tumor from patients with negative (pN0 and/or pM0) or positive (pN1 and/or pM1 and/or metachronous) metastatic status. *, one Ren-PDOX has not been evaluated for lung metastasis.

Overall mass architecture of paired human specimens and Ren-PDOX tumors observed by HE staining (20X). TNM staging and Fuhrman grade (F) of implanted tumors are reported. For Ren-PDOXs, lung micrometastases in HE and vimentin stained sections are shown (40X).

Human to mouse stroma substitution. CD34/CD31 staining for respectively human (h) and mouse (m) blood vessels in original human specimens and paired Ren13 and 28-PDOX are shown (20X). Ren-PDOX OT referred to tumor generated from first implantation of human specimen in mice. Insets show human and mouse kidney as positive/negative controls of staining (20X).

Frequency histogram of SNV and/or indels found in Ren-PDOX by high-throughput sequencing in Ren13, Ren28, Ren36, Ren86 (targeted-NGS analysis by Haloplex) and Ren50-PDOX (WES). Only non-synonymous/stop gain SNVs and frameshift indels novel or with MAF<0.01 are included.

Genetic comparison of original human specimen (patient #50) and its derived PDOX (Ren50-PDOX) by WES. Venn diagram shows number and percentage of variants called in human specimen (yellow), in its PDOX (blue) and overlapped variants (green).
Kidney Cancer PDOXs reveal patient-specific pro-malignant effects of antiangiogenics and its molecular traits (2020)
Figure 4

Figure 4. Effects of anti-VEGF/R therapies on local invasion.

Capsular invasion was defined as i) absent if tumor cells were separated from normal renal parenchyma by well-defined layers of FN or ii) present if strands of tumor cells were directly in contact with normal renal parenchyma

B,C) Effects of DC and Beva treatment on Ren13, Ren86, Ren28 and Ren50-PDOX on capsular invasion (B) and tumor front of invasion (C). (B) Incidence of Capsular invasion (stripes) in each model and treatment. 5-8 animals per model and treatment group were analyzed by Chi-square test where *p < 0.05, **p < 0.01. (C) Representative images of Vimentin staining (4X), as described in Figure 1, and box plot quantification of fold-invasion (∆ invasion) of treated vs control tumors indicating median, Q1/Q3 and max/min value whiskers. 6-12 animals per model and treatment group were analyzed by Mann-Whitney test where *p < 0.05, **p < 0.01.

D) Effects of sunitinib treatment on Ren13BM and Ren28 on capsular invasion and tumor front invasion. Bar graphs show incidence of capsular invasion (stripes) in control and sunitinib treated tumors. 3-9 animals per model and treatment group were analyzed by Chi-square test where *p < 0.05, **p < 0.01. Box plots represent median, Q1/Q3 and max/min value whiskers of fold-invasion (∆ invasion) of sunitinib-treated vs control tumors. 5-8 animals per model and treatment group were analyzed by Mann-Whitney test. *p < 0.05, **p < 0.01.