Figure 1. FN stimulates ERα’s... | Rockefeller University Press

Figure 1.

$Figure 1. FN stimulates ERα’s transcriptional activity. (a) Luciferase assay in MCF7 cells transiently transfected with pTK-ERE-Luc and pTK-Renilla, seeded on BSA or FN and treated for 14 h as indicated. Data are represented as mean ± SD. Differences between groups were analyzed by two-way ANOVA followed by Bonferroni contrasts adjusted for multiple comparisons (n = 3 replicates). (b and c) Top: Western blot of a subcellular fractionation of MCF7 cells seeded on BSA and treated for 15 min (b) or 60 min (c) as indicated. Blotting antibodies are shown on the left. Bottom: densitometry. For each subcellular fraction, shown is the ERα/β-actin density ratio normalized to the mean control group. Each symbol represents a different experiment. Differences between groups were analyzed by a one-tailed paired Student’s t test (n = 3 replicates). (d and e) Top: Western blot of a subcellular fractionation of MCF7 cells seeded on FN and treated for 15 (d) or 60 min (e) as indicated. Blotting antibodies are shown on the left. Bottom: Densitometry. For each subcellular fraction, shown is the ERα/β-actin density ratio normalized to the mean control group. Each symbol represents a different experiment. (f) Western blot of the cytosolic + membrane fraction of MCF7 cells seeded on BSA or FN and treated with E2 for the indicated times. Below the blots, the ERα/β-actin density ratio is shown, normalized to the control group. (g) Western blot of the nuclear fraction of MCF7 cells seeded on BSA or FN and treated with E2 for the indicated times. Below the blots, the ERα/PCNA density ratio is shown, normalized to the control group. Differences between groups were analyzed by one-tailed paired Student’s t test (n = 3 replicates). *, P < 0.05; **, P < 0.01; ***, P < 0.001. Shown data are representative of at least three independent experiments. Black arrowheads indicate positions of 50-kD markers. White arrowheads indicate positions of 37-kD markers. Treatments: ethanol (vehicle) or 10−8 M E2.$

FN stimulates ERα’s transcriptional activity. (a) Luciferase assay in MCF7 cells transiently transfected with pTK-ERE-Luc and pTK-Renilla, seeded on BSA or FN and treated for 14 h as indicated. Data are represented as mean ± SD. Differences between groups were analyzed by two-way ANOVA followed by Bonferroni contrasts adjusted for multiple comparisons (n = 3 replicates). (b and c) Top: Western blot of a subcellular fractionation of MCF7 cells seeded on BSA and treated for 15 min (b) or 60 min (c) as indicated. Blotting antibodies are shown on the left. Bottom: densitometry. For each subcellular fraction, shown is the ERα/β-actin density ratio normalized to the mean control group. Each symbol represents a different experiment. Differences between groups were analyzed by a one-tailed paired Student’s t test (n = 3 replicates). (d and e) Top: Western blot of a subcellular fractionation of MCF7 cells seeded on FN and treated for 15 (d) or 60 min (e) as indicated. Blotting antibodies are shown on the left. Bottom: Densitometry. For each subcellular fraction, shown is the ERα/β-actin density ratio normalized to the mean control group. Each symbol represents a different experiment. (f) Western blot of the cytosolic + membrane fraction of MCF7 cells seeded on BSA or FN and treated with E₂ for the indicated times. Below the blots, the ERα/β-actin density ratio is shown, normalized to the control group. (g) Western blot of the nuclear fraction of MCF7 cells seeded on BSA or FN and treated with E₂ for the indicated times. Below the blots, the ERα/PCNA density ratio is shown, normalized to the control group. Differences between groups were analyzed by one-tailed paired Student’s t test (n = 3 replicates). *, P < 0.05; **, P < 0.01; ***, P < 0.001. Shown data are representative of at least three independent experiments. Black arrowheads indicate positions of 50-kD markers. White arrowheads indicate positions of 37-kD markers. Treatments: ethanol (vehicle) or 10⁻⁸ M E₂.