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Fig. 8 | Journal of Experimental & Clinical Cancer Research

Fig. 8

From: RETRACTED ARTICLE: Receptor tyrosine kinase inhibitor Sunitinib and integrin antagonist peptide HM-3 show similar lipid raft dependent biphasic regulation of tumor angiogenesis and metastasis

Fig. 8

Distribution of integrin α5β1, αvβ3 and VEGFR2 on EAhy926 cells by HM-3 and Sunitinib treatment after cleavage of glypican-1. a Treatment strategy. Samples 1–6 are membrane fractions outside of the lipid raft region and samples 7–12 are membrane fractions inside the lipid raft region of the corresponding cells. b Detection of human transferrin R (non-raft marker) and caveolin-1 (raft marker) with Western-blot analysis. The distribution of glypican-1 was also analyzed. c Western-blot analysis to show redistribution of integrin α5β1 and αvβ3 by HM-3 and Sunitinib treatment after glypican-1 cleavage. Intensities of the protein bands were analyzed with Image J and shown as histograms in panel d for integrin α5β1 and panel e for integrin αvβ3. Statistical analysis was performed to samples 2 and 8. f Western-blot analysis to show distribution of VEGFR2 and p-VEGFR2 by HM-3 and Sunitinib treatment after glypican-1 cleavage. Intensities of the protein bands were shown as histograms in panel g for VEGFR2 and panel h for p-VEGFR2. Statistical analysis was performed to compare samples 2 and 8. i Immunoprecipitation was performed with anti-integrin αvβ3 antibodies and VEGFR2 or p-VEGFR2 was detected with Western-blot analysis to show distribution of integrin αvβ3-VEGFR2 complexes and the corresponding p-VEGFR2 on glypican-1-cleaved cells. Intensities of the protein bands were analyzed with Image J and shown as histograms in panel j for VEGFR2 and panel k for p-VEGFR2. Statistical analysis was performed comparing sample 2 for the non-raft region and sample 8 for the raft region. Data are represented as mean ± SD (*p<0.05, **p<0.01)

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