(A) Relative transforming growth factor beta induced (TGFBI) expression of mesenchymal transformed breast tumor cell lines compared to noninvasive settings was assessed using real-time quantitative PCR

(A) Relative transforming growth factor beta induced (TGFBI) expression of mesenchymal transformed breast tumor cell lines compared to noninvasive settings was assessed using real-time quantitative PCR. = 4; T47D-EMT = 3; *< 0.05 (E) Relative SNAI1 expression of mesenchymal transformed breast cancer cell lines compared to non-invasive controls was assessed using real-time quantitative PCR. Data symbolize imply SEM. Using unpaired, two-tailed = 4; T47D-EMT = 5; ***< 0.0005 (F) Relative SNAI2 expression of mesenchymal transformed breast cancer cell lines compared to non-invasive controls was assessed using real-time quantitative PCR. Data symbolize imply SEM. Using unpaired, two-tailed = 3; T47D-EMT = 4; *< 0.05. Demonstration_1.zip (478K) GUID:?B9997237-57D4-47A1-8944-691144FBA691 Number S2: CYR61 expression correlates with breast tumor cell invasiveness. (A) Relative CYR61 manifestation 96 h after transient CYR61 siRNA transfection compared to control was assessed using real-time quantitative PCR. Data symbolize imply SEM. Using unpaired, two-tailed = 8; T47D-EMT = 7; MDA-MB-231 = 3; HCC1806 = 4; ** < 0.01; ***< 0.0001 (B) Analysis of family member AlamarBlue reduction as indication for cell viability. Transient transfected breast tumor cell spheroids were cultivated and AlamarBlue reduction was assessed 48 hours after adding Matrigel at 4 h incubation. Relative AlamarBlue reduction was calculated compared to control spheroids. Data symbolize imply SEM. = 3 (C) Analysis of relative AlamarBlue reduction as indication for cell viability. Breast tumor cell spheroids were cultivated and AlamarBlue reduction was assessed 48 h after adding Matrigel and 1g/ml rhCYR61 at 4 h incubation. Relative AlamarBlue reduction was calculated compared to control spheroids. Data symbolize imply SEM. = 3. Demonstration_1.zip (478K) GUID:?B9997237-57D4-47A1-8944-691144FBA691 Number S3: Suppression of CYR61 reduces S100A4 expression. MC-GGFG-DX8951 (A) Immunoblot analysis of S100A4 mRNA manifestation levels in different breast tumor cell lines 96 h after S100A4 siRNA transfection was recognized using western blotting. Date symbolize imply SEM. Using MC-GGFG-DX8951 unpaired, two-tailed = 4; T47D-EMT = 4; MDA-MB-231 = 3; HCC1806 = 3; *< 0.05< 0.01< 0.005 (B) Representative experiments of S100A4 protein expression quantification corresponding to (A). (C) S100A4 mRNA manifestation analysis 96 h after siRNA transfection using quantitative PCR. Day symbolize imply SEM. Using unpaired, two-tailed = 4; T47D-EMT = 4; MDA-MB-231 = 3; HCC1806 = 3; ***< 0.005; ****< 0.0001 (D) CYR61 mRNA expression analysis 96 h after S100A4 siRNA transfection using quantitative PCR. Day symbolize imply SEM. MC-GGFG-DX8951 MCF-7-EMT = 5; T47D-EMT = 6; MDA-MB-231 = 3; HCC1806 = 3 (E) Analysis of relative AlamarBlue reduction as indication for cell viability. Breast tumor cell spheroids transient transfected with S100A4 siRNA were cultivated and AlamarBlue reduction was assessed 48 h after adding Matrigel at 4 h incubation. Relative AlamarBlue reduction was calculated compared to control spheroids. Data symbolize imply SEM. = 3. Demonstration_1.zip (478K) GUID:?B9997237-57D4-47A1-8944-691144FBA691 Number S4: ERK1/2 activity is transducer of CYR61 mediated S100A4 regulation. (A) ERK1/2 and p-Erk1/2 (Thr202/Tyr204) manifestation in different breast tumor cell lines with or without 10M U0126 treatment recognized by western blotting. (B) ERK1/2 and p-Erk1/2 (Thr202/Tyr204) manifestation in noninvasive breast tumor cell lines with or without 1g/ml rhCYR61 treatment recognized by western blotting. Demonstration_1.zip (478K) GUID:?B9997237-57D4-47A1-8944-691144FBA691 Number S5: Suppression of YAP reduces invasiveness through blocking CYR61-S100A4-pERK1/2 signaling. (A) Relative YAP manifestation 96 h after transient YAP siRNA transfection compared to control was assessed using real-time quantitative PCR. Data symbolize imply SEM. Using unpaired, two-tailed = 5; T47D-EMT = 3; MDA-MB-231 = 3; HCC1806 = 3; *< 0.05< 0.01< 0.001 (B) Analysis of family member AlamarBlue reduction as HOXA2 indication for cell viability. Breast tumor cell spheroids were cultivated and AlamarBlue reduction was assessed.