Figure 4: Loss of MITF is accompanied by increased invasiveness.
From: Low MITF/AXL ratio predicts early resistance to multiple targeted drugs in melanoma
(a) PLX4720-resistant melanoma cells and their treatment-naïve counterparts were seeded on a matrigel-coated invasion chamber for 12 h. Cells that invaded through the matrix were stained with crystal violet. Scale bars, 500 μm. (b) Quantification of a from at least five images. (c) PLX4720-resistant cells (R) and their treatment-naïve (S) counterparts were blotted for EMT markers. α-Tubulin served as a loading control. (d) ZEB1 and ZEB2 mRNA were measured in acquired PLX4720-resistant cells and their treatment-naïve counterparts. The results are based on three independent experiments and normalized to RPL13. Data are shown as mean+s.d. (e) mRNA expression of Dia1 was measured in acquired PLX4720-resistant cells and their treatment-naïve counterparts. The results are based on three independent experiments and normalized to RPL13. Data are shown as mean +s.d. (f) Knockdown of Fra-1 in two PLX4720-resistant cell lines was documented on immunoblot. HSP90 served as a loading control. (g) Two PLX4720-resistant melanoma cells transduced with Fra-1 knockdown and control cells were seeded on a matrigel-coated invasion chamber for 12 h. Cells that invaded through the matrix were stained with crystal violet. Inserts show proliferation in 12 h on polystyrene culture dishes. Scale bars, 200 μm. (h) Quantification of g of at least five images.
