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. 2017 Nov 27;8(66):110166–110175. doi: 10.18632/oncotarget.22698

Figure 2. ID3 regulates cell migration but not cell proliferation or cell cycle states and it regulates SOX10/ MITF expression.

Figure 2

(A) Migration rate of ID3-engineered cell lines (A375, SKmel28, HT144 and WM266-4) was measured as a percentage of the gap closure at the indicated time points. Bottom pictures show representative images for each condition. (B) Cell proliferation of ID3-engineered cell lines (A375, SKmel28, HT144 and WM266-4) was measured by Alamar blue staining everyday up to 5 days. Relative Fluorescence Units (RFU) are represented in 103 values. (C) Analysis of cell cycle stages by flow cytometry after PI staining of ID3-engineered cell lines (A375, SKmel28 and WM266-4). (D) Quantitative real-time PCR analysis of mesenchymal markers FN1, SNAIL and SLUG in ID3 knockdown cell lines (A375, SKmel28 and HT144). (E) Quantitative real-time PCR analysis of MITF and SOX10 expression in ID3 knockdown melanoma cell lines (A375, SKmel28 and HT144). (F) Quantitative real-time PCR analysis of SOX10 in ID3 overexpressing WM266-4 cell line. A. – E.: Data represent mean ± SD of biological triplicates. *P < 0.05, **P < 0.01, ***P < 0.001.