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. 2018 Dec 3;7:e27706. doi: 10.7554/eLife.27706

Figure 5. SOX4 controls tumor-induced angiogenesis in vivo in a zebrafish tumor-xenograft model.

(A) Schematic representation of the zebrafish tumor-xenograft model, with the subintestinal vessels indicated in green (SIV) and tumor cells in red. (B) Representative images of zebrafish injected with ER-control and ERSOX4 HMLE cells treated with 4-OHT. Arrows indicate newly formed blood vessels. (C) Quantification of the number of ectopic sprouts observed per fish injected with ER-control and ERSOX4 HMLE cells. Individual data points and data represented as mean ± SD of three independent biological replicates (**p-value<0.01, ***p-value<0.001; ANOVA). (D) Quantification of the relative increase in ectopic sprouting in fish injected with 4-OHT treated relative to untreated ER-control and ERSOX4 cells. Data represented as mean ± SD of three independent biological replicates (* p-value<0.05, Student’s t-test). (E) Quantification of the number of ectopic sprouts in fish injected with SCR control and SOX4 KD MDA-MB-231 cells. Results obtained from three independent biological replicates (***p-value<0.001, Student’s t-test). (F) Representative images of zebrafish injected with SCR control or SOX4 KD MDA-MB-231 cells. Arrows indicate newly formed blood vessels (G) Quantification of the number of fish injected with MDA-MB-231 cells with or without ectopic sprouting. Results obtained from three independent biological replicates (***p-value<0.001, Student’s t-test). (H) Quantification of the number of ectopic sprouts observed per fish injected with 4-OHT-treated ERSOX4 HMLE cells exposed with siRNA control or siRNA targeting ET-1. Individual data points and data represented as mean ± SD (*p-value<0.05, ANOVA). (I) Quantification of the relative increase in ectopic sprouting in fish injected with 4-OHT treated ERSOX4 HMLE cells exposed with siRNA control or siRNA targeting ET-1. Data represented as mean ± SD of three independent biological replicates (*p-value<0.05, ***p-value<0.001; ANOVA). (J) Representative images of zebrafish injected with 4-OHT-treated ERSOX4 HMLE cells exposed with siRNA control or siRNA targeting ET-1. Arrows indicate newly formed blood vessels.

Figure 5.

Figure 5—figure supplement 1. Analysis of angiogenic potential of HMLE WT cells.

Figure 5—figure supplement 1.

SOX4 and EDN1 mRNA expression in SOX4-depleted MDA-MB-231 cells. (A) Quantification of ectopic sprouting in zebrafish tumor-xenograft model using wild-type (WT) HMLE cells in presence of absence of 4-OHT. (B) qPCR validation of SOX4 knockdown (KD) relative to SCR control in MDA-MB-231 cells with matching EDN1 expression.