Fig. 5
Modification of SUSD2 levels and detection of SUSD2 in CTC of breast cancer patients.A: Downregulation of the SUSD2 protein in CTC-ITB-01 by siRNA, analyses by Western Blot.B: Quantitative Western Blot analysis of the PDCD4 and GRP78 levels shown in (A).C: Response of SUSD2 and PDCD4 to 14 days of hypoxia (1% O2; 14 d) and reoxygenation (14 days of hypoxia, followed by 10% O2 for 4 h; + 4 h). ERα (MCF-7) and EGFR (MDA-MB-468) were analyzed as an established reaction to hypoxia. S: standard cell culture condition.D: Quantitative Western Blot analysis of the SUSD2 and PDCD4 levels shown in (C).E: Detection of SUSD2 in CTC of breast cancer patients using the CellSearch system.F: Isolation of SUSD2 positive CTCs from the blood of breast cancer patients by MACS. Slides were stained with anti-mouse IgG to detect the MicroBeads bound tumor cells (IgG). The top row shows a SUSD2 positive CTC cluster isolated by MACS. The middle row shows an overview of an elution fraction on a cytospin. The SUSD2 positive CTCs are labelled with the white box and are enlarged in the bottom row.A-D,F: All experiments were performed in biological triplicates. B, D: Average values are shown as numbers, and the standard deviation as vertical error bars. The p-values were calculated using Student’s two-sided t-test where p < 0.05 was considered significant.E: The CellSearch Results are specified in Supplementary Table S5 (E). Uncropped X-ray film images are shown in supplementary file F1.
