Gene expression of the A375 melanoma cell line treated with RAF inhibitors Vemurafenib and time-course with EGF stimulation during Vemurafenib RAF inhibitor treatment alone or with the MEK inhibitor Cobimetinib. Gene expression of the A375 melanoma cell line treated with RAF inhibitors Vemurafenib and time-course with EGF stimulation during Vemurafenib RAF inhibitor treatment alone or with the MEK inhibitor Cobimetinib

We treated for 24 hours the BRAF-V600E melanoma cell line A375 with 7 doses of the RAF inhibitor Vemurafenib and, in a second experimental desing, we treated for 24 hours the BRAF-V600E melanoma cell line A375 with Vemurafenib (1 uM) alone or in combination with the MEK inhibitor Cobimetinib (1 uM) and subsequently stimulated with EGF in a time-course of 7 time points for up to 8 hours (0, 0.5, 1, 2, 3, 4, 8 hours). Overall design: We measured mRNA levels of the A375 cell line treated for 24 hours with 7 doses of the RAF inhibitor Vemurafenib in the range 0.001-10 uM and, in a second experimental design, mRNA levels of the A375 cell line treated for 24 hours with the RAF inhibitor Vemurafenib (1 uM) alone or in combination with the MEK inhibitor Cobimetinib (1 uM) and subsequently stimulated with EGF (100 ng/mL) in a time-course up to 8 hours

我们将携带BRAF-V600E突变的黑色素瘤细胞系A375以7个梯度剂量的RAF抑制剂（RAF inhibitor）维莫非尼（Vemurafenib）处理24小时；在第二项实验设计中，我们同样对A375细胞采用1 μM维莫非尼单独处理，或联合使用1 μM的MEK抑制剂（MEK inhibitor）考比替尼（Cobimetinib）处理24小时，随后用表皮生长因子（EGF）进行最长8小时的时间梯度刺激，共设置7个检测时间点：0、0.5、1、2、3、4、8小时。整体实验设计如下：我们检测了经0.001~10 μM梯度剂量的RAF抑制剂维莫非尼处理24小时的A375细胞的mRNA水平；在第二项实验设计中，我们检测了经以下处理的A375细胞的mRNA水平：单独使用1 μM维莫非尼处理24小时，或联合使用1 μM的MEK抑制剂考比替尼处理24小时，随后以100 ng/mL的表皮生长因子进行最长8小时的时间梯度刺激。