serpine2 protein  (MedChemExpress)

Journal: Oncogene

Article Title: ZEB1-repressed microRNAs inhibit autocrine signaling that promotes vascular mimicry of breast cancer cells

doi: 10.1038/onc.2017.356

Figure Lengend Snippet: COPII-mediated secretion is essential for VM. ( a ) Quantitative reverse transcription–PCR (qRT–PCR) for SEC13 in MDA-MB-231 or BT-549 cells transfected with siNT or siSEC13. Error bars represent s.d. of three (MDA-MB-231) or two (BT-549) independent experiments. ( b ) Representative images of VM in MDA-MB-231 or BT-549 cells transfected with siNT or siSEC13. Scale bars=1000 μm. ( c ) Quantification of total network length in VM assays in MDA-MB-231 or BT-549 cells. Error bars represent s.d. for five or three independent experiments, respectively. T -tests were performed to compare each to siNT control, *** P <0.001, * P <0.05. ( d ) qRT–PCR following expression of a NEG or clustered miRNA mimics. Error bars represent s.d. of three independent experiments. T -tests were performed vs NEG control, *** P <0.001. ( h ) Western blot for FN1 and SERPINE2 in conditioned media from MDA-MB-231 cells transfected as shown. ( f ) Quantification of three (FN1) or two (SERPINE2) independent replicates of western blots as shown in e . Error bars represent s.d. T -tests were performed for FN1 quantitation compared to NEG control. ** P <0.05; *** P <0.001. ( g ) qRT–PCR following transfection of MDA-MB-231 cells with miRNA mimics. Error bars represent s.d. from three independent experiments. T -tests were performed, * P <0.05, ** P <0.01, *** P <0.001. ( h ) Western blot for LRP1 following transfection of MDA-MB-231 cells with miRNA mimics. Shown is a representative image of two independent experiments. GAPDH is loading control. ( i ) qRT–PCR following transfection of MDA-MB-231 cells with miRNA mimics. Error bars represent s.d. from three independent experiments. T -tests were performed, * P <0.05, *** P <0.001.

Article Snippet: Recombinant FN1 (1918-FN-02M, R&D Systems, Minneapolis, MN, USA) and SERPINE2 (2980-PI-010, R&D Systems) were added to VM assays at concentrations of 1 and 10 μg/ml, respectively.

Techniques: Reverse Transcription, Quantitative RT-PCR, Transfection, Control, Expressing, Western Blot, Quantitation Assay

Journal: Oncogene

Article Title: ZEB1-repressed microRNAs inhibit autocrine signaling that promotes vascular mimicry of breast cancer cells

doi: 10.1038/onc.2017.356

Figure Lengend Snippet: Increased expression of FN1 and SERPINE2 is regulated by serum withdrawal and is essential for VM. ( a ) Quantitative reverse transcription–PCR (qRT–PCR) in breast cancer cell lines cultured with 10 or 0% serum for 48 h. Error bars represent s.d. of three independent experiments. T -tests were performed for each line comparing 0vs10% conditions, ** P <0.01, *** P <0.001. ( b , d ) Representative images of VM in control and FN1 ( b ) or SERPINE2 ( d ) knockdown cells. Scale bars=1000 μm. ( c , e ) Quantification of total network length for VM assays as shown in b and d . Error bars represent s.d. for at five and four independent experiments, respectively. T -tests were performed to compare each to siNT control, ** P <0.01. ( f ) Representative images of VM in control and FN1 or SERPINE2 knockdown in BT-549 cells. Scale bars=1000 μm. ( g ) Quantification of total network length. Error bars represent s.d. for three independent experiments. T -tests were performed to compare each to siNT control, * P <0.05. ( h ) Representative phase images of cell morphology in siNT or siSERPINE2 cells. Scale bars=100 μm. ( i ) qRT–PCR for FN1 and CDH1 in cells transfected with siNT or siSERPINE2. Error bars represent s.d. of three independent experiments. T -test was performed, *** P <0.001. ( j ) qRT–PCR for FN1 and SERPINE2 in cells transfected with NEG or clustered miRNA mimics for 48 h, then cultured with 10 or 0.5% serum for 48 h. Error bars represent s.d. of three independent experiments. T -tests were performed comparing 0.5vs10% in the control or comparing cells transfected with miRNAs vs NEG in matching serum conditions, *** P <0.001. ( k ) Western blot for FN1 for cells as in h . ACTB is loading control. ( l , m ) Quantification of total network length of VM assays from VM-incompetent cells plated in 0% serum ( l ) or VM-competent cells plated in 5% serum ( m ) in the presence or absence of recombinant FN1 and SERPINE2 as shown. Error bars represent s.d. of two independent experiments.

Article Snippet: Recombinant FN1 (1918-FN-02M, R&D Systems, Minneapolis, MN, USA) and SERPINE2 (2980-PI-010, R&D Systems) were added to VM assays at concentrations of 1 and 10 μg/ml, respectively.

Techniques: Expressing, Reverse Transcription, Quantitative RT-PCR, Cell Culture, Control, Knockdown, Transfection, Western Blot, Recombinant

Journal: Oncogene

Article Title: ZEB1-repressed microRNAs inhibit autocrine signaling that promotes vascular mimicry of breast cancer cells

doi: 10.1038/onc.2017.356

Figure Lengend Snippet: Autocrine signaling factors FN1, ITGB1, SERPINE2 and LRP1 predict poor survival in early-stage breast cancer. ( a ) Plots comparing FN1 , ITGB1 , SERPINE2 and LRP1 levels in RNA-seq data from subtyped breast cancer cell lines. A one-way analysis of variance with Tukey’s test was performed, * P <0.5, *** P <0.001. ( b – f ) Survival curves from cBioPortal showing survival of patients with stage I, II or III breast cancer with the following genes altered. Log-rank test P -values are shown. ( b ) Tumors that had a low level (defined by heterozygous loss or homozygous deletion) of MIR200C along with high expression (defined by a gain, amplification or fold expression >2) of FN1 and SERPINE2 (red) compared to those that did not (blue). ( c ) Tumors that had a low level of MIR183 along with high expression of FN1 and SERPINE2 (red) compared to those that did not (blue). ( d ) Tumors that had a high level of FN1 , ITGB1 , SERPINE2 and LRP1 (red) compared to those that did not (blue). ( e ) Tumors that had a high level of FN1 and ITGB1 (red) compared to those that did not (blue). ( f ) Tumors that had a high level of SERPINE2 and LRP1 (red) compared to those that did not (blue).

Article Snippet: Recombinant FN1 (1918-FN-02M, R&D Systems, Minneapolis, MN, USA) and SERPINE2 (2980-PI-010, R&D Systems) were added to VM assays at concentrations of 1 and 10 μg/ml, respectively.

Techniques: RNA Sequencing, Expressing, Amplification