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β- induced MCF10A cells. A. mRNA expression levels of DLC1, SNAIL1, and <t>ZEB1</t> at early (2-day) and late (5-day) EMT time points measured by QPCR. Data represents the mean+SD of four biological replicates. B. Protein expression levels measured by Western blot. Quantified fold change for three biological replicates. One-way ANOVA was performed for untreated-TGFβ comparisons with Sidak’s multiple comparison test (ns not significant, *P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001). The p-values, including the additional siCtrl-siDLC1 comparisons, were listed in .
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β- induced MCF10A cells. A. mRNA expression levels of DLC1, SNAIL1, and <t>ZEB1</t> at early (2-day) and late (5-day) EMT time points measured by QPCR. Data represents the mean+SD of four biological replicates. B. Protein expression levels measured by Western blot. Quantified fold change for three biological replicates. One-way ANOVA was performed for untreated-TGFβ comparisons with Sidak’s multiple comparison test (ns not significant, *P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001). The p-values, including the additional siCtrl-siDLC1 comparisons, were listed in .
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β- induced MCF10A cells. A. mRNA expression levels of DLC1, SNAIL1, and ZEB1 at early (2-day) and late (5-day) EMT time points measured by QPCR. Data represents the mean+SD of four biological replicates. B. Protein expression levels measured by Western blot. Quantified fold change for three biological replicates. One-way ANOVA was performed for untreated-TGFβ comparisons with Sidak’s multiple comparison test (ns not significant, *P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001). The p-values, including the additional siCtrl-siDLC1 comparisons, were listed in .

Journal: PLOS Computational Biology

Article Title: Integrated mathematical and experimental modeling uncovers enhanced EMT plasticity upon loss of the DLC1 tumor suppressor

doi: 10.1371/journal.pcbi.1013076

Figure Lengend Snippet: β- induced MCF10A cells. A. mRNA expression levels of DLC1, SNAIL1, and ZEB1 at early (2-day) and late (5-day) EMT time points measured by QPCR. Data represents the mean+SD of four biological replicates. B. Protein expression levels measured by Western blot. Quantified fold change for three biological replicates. One-way ANOVA was performed for untreated-TGFβ comparisons with Sidak’s multiple comparison test (ns not significant, *P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001). The p-values, including the additional siCtrl-siDLC1 comparisons, were listed in .

Article Snippet: The membranes incubated with primary antibodies (anti-DLC1 mouse mAb (1:500, BD Biosciences Cat# 612021, RRID:AB_399416), anti-SNAIL1 rabbit mAb (1:1000, Cell Signaling Technology Cat# C15D3), anti-ZEB1 rabbit mAb (1:1000, Cell Signaling Technology Cat# D80D3), anti-ACTB mouse mAb (1:2000, Sigma-Aldrich Cat# A1978, RRID:AB_476692), anti-pERK-p44/42 rabbit pAb (1:1000, Cell Signaling Technology Cat# 9101, RRID:AB_331646) in blocking solution supplied with 0.05% sodium azide overnight and was followed by HRP-labeled secondary antibody incubation for 1h.

Techniques: Expressing, Western Blot, Comparison

A. Interaction graph of the CBSD model. Interactions of the original CBS model are indicated with black arrows. Additional regulations through the integration of DLC1 are marked with red arrows. B. Fit of the CBSD model trajectories (solid lines) to the CBS model’s simulated data (stars). dlc1 data in the siCtrl was included relative to the zeb1 simulated data. The model was equilibrated without exogenous TGFβ stimulation for 100 timesteps and afterward stimulated with 10 ng/ml exogenous TGFβ. C. Fit of the CBSD model to the experimental data under dlc1 knockdown. The stars represent the mean observed fold-changes of dlc1 and snail1 upon dlc1 knockdown at day 5 , and the lines represent the respective model simulation values. D. Exogenous TGFβ (TGF0) versus N-cad bifurcation of the CBSD model under control condition (blue line) compared to the bifurcation diagram of the original CBS model (grey line).

Journal: PLOS Computational Biology

Article Title: Integrated mathematical and experimental modeling uncovers enhanced EMT plasticity upon loss of the DLC1 tumor suppressor

doi: 10.1371/journal.pcbi.1013076

Figure Lengend Snippet: A. Interaction graph of the CBSD model. Interactions of the original CBS model are indicated with black arrows. Additional regulations through the integration of DLC1 are marked with red arrows. B. Fit of the CBSD model trajectories (solid lines) to the CBS model’s simulated data (stars). dlc1 data in the siCtrl was included relative to the zeb1 simulated data. The model was equilibrated without exogenous TGFβ stimulation for 100 timesteps and afterward stimulated with 10 ng/ml exogenous TGFβ. C. Fit of the CBSD model to the experimental data under dlc1 knockdown. The stars represent the mean observed fold-changes of dlc1 and snail1 upon dlc1 knockdown at day 5 , and the lines represent the respective model simulation values. D. Exogenous TGFβ (TGF0) versus N-cad bifurcation of the CBSD model under control condition (blue line) compared to the bifurcation diagram of the original CBS model (grey line).

Article Snippet: The membranes incubated with primary antibodies (anti-DLC1 mouse mAb (1:500, BD Biosciences Cat# 612021, RRID:AB_399416), anti-SNAIL1 rabbit mAb (1:1000, Cell Signaling Technology Cat# C15D3), anti-ZEB1 rabbit mAb (1:1000, Cell Signaling Technology Cat# D80D3), anti-ACTB mouse mAb (1:2000, Sigma-Aldrich Cat# A1978, RRID:AB_476692), anti-pERK-p44/42 rabbit pAb (1:1000, Cell Signaling Technology Cat# 9101, RRID:AB_331646) in blocking solution supplied with 0.05% sodium azide overnight and was followed by HRP-labeled secondary antibody incubation for 1h.

Techniques: Knockdown, Control