Data files for figures 1-8

Data file 1. Data for figures 1A, B and D and data not shown.A, B) Relative mRNA expression from the indicated cell lines is shown for three replicates from independent mRNA isolations. The numbers reflect qPCR delta Ct numbers using 18S rRNA Ct values. The values were scaled to an average of 1.0 for Scp-2 cells. The primers used are shown in Materials and Methods.D) Relative MMP-1 protein expression. Three immunoblots were performed with anti-MMP1 sera and lysates of the indicated cell lines as in figure 1C. The MMP-1 band intensities were quantified using the Odyssey infrared imager (Li-Cor) and the software provided by the manufacturer. The intensities were scaled to the average intensities of MMP-1 in Scp-2 cells.Data not shown) Relative MMP-1 expression in other MDA-MB-231 single cell population cell lines as in figure 1A. Values are from two independent replicates.
Data file 2. Data for figures 2C and E.Relative luciferase activity is shown for three replicates. Each replicate is the average of duplicate determinations (i.e. six total points). The firefly luciferase values were divided by the Renilla luciferase values (from cotransfected plasmid pRLSV40P). In (C) the values were scaled to an average of 1.0 for the Scp-2 cells and the -819 to +71 MMP1 reporter, except for CMV which was scaled to its own values for Scp-2 cells. In (E) the values were scaled in each replicate to 1.0 for Scp-2 cells and the -172/-27 MMP1 reporter.
Data file 3. Data for figures 3B and D.Relative luciferase activity is shown for three replicates. Each replicate is the average of duplicate determinations (i.e. six total points). The firefly luciferase values were divided by the Renilla luciferase values (from cotransfected plasmid pRLSV40P). The values were scaled in each replicate to 1.0 for Scp-2 cells and the wild type reporter. For the synthetic reporter, the values were scaled to that of the 3X-AP1 site reporter.
Data file 4. Data for figures 4A and C.A) Relative mRNA expression from the indicated cell lines and genes is shown for three replicates from independent mRNA isolations. The numbers reflect qPCR Ct numbers using 18S rRNA Ct values. The values were scaled to a value of 1.0 for Scp-2 cells. The primers used are shown in Materials and Methods.C) Relative Fra-1 protein expression. Three immunoblots were performed with anti-Fra-1 sera and lysates of the indicated cell lines as in figure 4B. The Fra-1 band intensities were quantified using the Odyssey infrared imager (Li-Cor) and the software provided by the manufacturer. The intensities were scaled to the average values for Fra-1 protein in Scp-2 cells.
Data file 5. Data for figures 5A, C and D.A) Relative Fra-1 mRNA expression in Scp-2 cells treated with the indicated siRNAs. Expression determined by qPCR is shown for three replicates from independent mRNA isolations. The numbers reflect qPCR delta Ct numbers using 18S rRNA Ct values. The values were scaled to an average value of 1.0 for untreated Scp-2 cells. The primers used are shown in Materials and Methods.B) Relative Fra-1 protein expression. Three immunoblots were performed with anti-Fra-1 sera and lysates of Scp-2 cells treated with the indicated siRNAs as in figure 5B. The Fra-1 band intensities were quantified using the Odyssey infrared imager (Li-Cor) and the software provided by the manufacturer. The values were scaled to the average intensities of Fra-1 protein in Scp-2 cells.C) As in (A) exept that MMP-1 and GAPDH mRNA expression were measured by qPCR. The primers used are shown in materials and methods.
Data file 6. Data for figure 6C.Chromatin immunoprecipitation from the indicated cell lines and using anti-Fra1 or control (no antibody) was quantified by qPCR using the indicated primers. The binding values were divided by those of input DNA and are shown as percent of input. Values from three replicate chromatin immunoprecipitations are shown.
Data file 7. Data for figures 7B and D.B) Scp-2 or Scp-21 cells were treated with the protein synthesis inhibitor cycloheximide for the indictate times. The cell lysates were then immunoblotted with anti-Fra-1 antibodies as in figure 7A. The Fra-1 protein band intensities were quantified on the Odyssey infrared imager (Li-Cor). The values for three independent experiments are shown. The values were scaled to 1.0 for cells treated for 0 hours (i.e. untreated).D) Scp-2 and Scp-21 cells were metabolically labeled with 35S-methionine and –cysteine for the indicated times. Fra-1 protein was then immunoprecipitated with anti-Fra-1 antibodies, run on SDS-PAGE and exposed to film. The bands were quantified using ImageJ software. The values were scaled to the value of Fra-1 expression in Scp-21 cells at 15 minutes of labeling. Values for there independent replicates are shown.
Data file 8. Data for figures 8A, D and E.A) Relative MMP-1 mRNA expression in the indicated cell lines with no expression vector, control pBabepuro vector or Fra-1 expressing pBabFra-1 vector. Expression determined by qPCR is shown for three replicates from independent mRNA isolations. The numbers reflect qPCR delta Ct numbers using 18S rRNA Ct values. The values were scaled to an average value of 1.0 for untreated Scp-2 cells. The primers used are shown in Materials and Methods.D) Quantfication of cell motility as in Fig. 8C. The number of cells crossing the initial scratch after 18 hours were counted in three independent experiments.E) Quantification of anchorage independent growth in soft agar. The number of colonies growing after 21 days of plating in soft agar were counted in three independent experiments.

数据文件1：包含图1A、B和D以及未展示数据的实验数据。A、B）展示特定细胞系中相对mRNA表达水平，数据来源于独立进行的mRNA分离的三个重复实验。数值反映了使用18S rRNA Ct值计算出的qPCR delta Ct数。数值已按Scp-2细胞的平均值调整为1.0。所使用的引物在材料和方法中展示。D）相对MMP-1蛋白表达。使用抗-MMP1血清和指示细胞系的裂解物进行三个免疫印迹实验，如图1C所示。使用Odyssey红外成像仪（Li-Cor）和制造商提供的软件对MMP-1条带强度进行量化。强度已按Scp-2细胞中MMP-1的平均强度进行调整。未展示数据）其他MDA-MB-231单细胞种群细胞系中相对MMP-1表达，如图1A所示。数值来自两个独立的重复实验。
数据文件2：包含图2C和E的实验数据。展示相对荧光素酶活性，数据来源于三个重复实验。每个重复实验是两次测定的平均值（即总共六个数据点）。萤火虫荧光素酶值除以Renilla荧光素酶值（来自共转染的质粒pRLSV40P）。在（C）中，数值已按Scp-2细胞和-819至+71 MMP1报告基因的平均值调整为1.0，CMV除外，其数值按Scp-2细胞的自身值进行调整。在（E）中，每个重复实验中的数值均按Scp-2细胞和-172/-27 MMP1报告基因的平均值调整为1.0。
数据文件3：包含图3B和D的实验数据。展示相对荧光素酶活性，数据来源于三个重复实验。每个重复实验是两次测定的平均值（即总共六个数据点）。萤火虫荧光素酶值除以Renilla荧光素酶值（来自共转染的质粒pRLSV40P）。每个重复实验中的数值均按Scp-2细胞和野生型报告基因的平均值调整为1.0。对于合成报告基因，数值调整为3X-AP1位点的报告基因。
数据文件4：包含图4A和C的实验数据。A）展示特定细胞系和基因的相对mRNA表达水平，数据来源于独立进行的mRNA分离的三个重复实验。数值反映了使用18S rRNA Ct值计算出的qPCR delta Ct数。数值已按Scp-2细胞的平均值调整为1.0。所使用的引物在材料和方法中展示。C）相对Fra-1蛋白表达。使用抗-Fra-1血清和指示细胞系的裂解物进行三个免疫印迹实验，如图4B所示。使用Odyssey红外成像仪（Li-Cor）和制造商提供的软件对Fra-1条带强度进行量化。强度已按Scp-2细胞中Fra-1蛋白的平均值进行调整。
数据文件5：包含图5A、C和D的实验数据。A）展示Scp-2细胞在处理指定siRNA后的相对Fra-1 mRNA表达水平。表达水平由qPCR确定，数据来源于独立进行的mRNA分离的三个重复实验。数值反映了使用18S rRNA Ct值计算出的qPCR delta Ct数。数值已按未处理Scp-2细胞的平均值调整为1.0。所使用的引物在材料和方法中展示。B）相对Fra-1蛋白表达。使用抗-Fra-1血清和Scp-2细胞处理指定siRNA的裂解物进行三个免疫印迹实验，如图5B所示。使用Odyssey红外成像仪（Li-Cor）和制造商提供的软件对Fra-1条带强度进行量化。数值已按Scp-2细胞中Fra-1蛋白的平均强度进行调整。C）与（A）类似，除了使用qPCR测量MMP-1和GAPDH mRNA表达。
数据文件6：包含图6C的实验数据。对指定细胞系进行染色质免疫沉淀，使用抗-Fra1或对照（无抗体），并通过qPCR使用指定的引物进行量化。结合值除以输入DNA的值，并显示为输入百分比。展示三个重复染色质免疫沉淀的数值。
数据文件7：包含图7B和D的实验数据。B）Scp-2或Scp-21细胞在指定时间内用蛋白质合成抑制剂环己酰亚胺处理。然后使用与图7A相同的抗-Fra-1抗体进行免疫印迹。在Odyssey红外成像仪（Li-Cor）上量化Fra-1蛋白条带强度。展示三个独立实验的数值。数值已按0小时处理（即未处理）的细胞进行调整。D）Scp-2和Scp-21细胞在指定时间内用35S-甲硫氨酸和-半胱氨酸进行代谢标记。然后使用抗-Fra-1抗体进行Fra-1蛋白的免疫沉淀，进行SDS-PAGE，并曝光。使用ImageJ软件量化条带。数值已按标记15分钟的Scp-21细胞中Fra-1表达值进行调整。展示三个独立重复的数值。
数据文件8：包含图8A、D和E的实验数据。A）展示指示细胞系中无表达载体、控制pBabepuro载体或Fra-1表达pBabFra-1载体下相对MMP-1 mRNA表达水平。表达水平由qPCR确定，数据来源于独立进行的mRNA分离的三个重复实验。数值反映了使用18S rRNA Ct值计算出的qPCR delta Ct数。数值已按未处理Scp-2细胞的平均值调整为1.0。所使用的引物在材料和方法中展示。D）与图8C类似的细胞迁移量化。在三个独立实验中计数18小时后穿过初始划痕的细胞数量。E）在软琼脂中独立生长的量化。在软琼脂中培养21天后计数生长的集落数量，在三个独立实验中进行。