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Bioss rabbit anti alox5 antibody
Rabbit Anti Alox5 Antibody, supplied by Bioss, used in various techniques. Bioz Stars score: 93/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+anti+gdf15+mic+polyclonal+antibody/GDF15+MIC-1+Polyclonal+Antibody/ppr0712718-96-18-22
Average 93 stars, based on 12 article reviews

rabbit anti alox5 antibody - by Bioz Stars, 2026-09

93/100 stars

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Western Blot:

Article Title: Increased plasma VEGF levels following ischemic preconditioning are associated with downregulation of miRNA-762 and miR-3072-5p.
Article Snippet: Cells were re-suspended in cold buffer containing 10-mM Tris-HCl (pH 7.4), 0.5-mM EGTA, 0.5-mM EDTA, 1% Triton X-100, and cOmpleteTM Mini Protease Inhibitor Cocktail (Sigma, St Louis, MO, USA) and were incubated for 30 min on ice and then centrifuged at 12 000 g for 20 min at 4 °C. .. Supernatants containing total protein were collected and subjected to Western blotting using rabbit anti-GDF15/ MIC-polyclonal antibody (1:500 dilution, # bs-3818R, Bioss, Woburn, Massachusetts, USA), anti-rabbit IgG HRP-conjugated secondary antibody (1:5000 dilution, #P0448, Dako, Glostrup, Denmark), and anti-β -actin HRP-conjugated antibody (1:2000 dilution #NB600-532H, Novus Biologicals, Littleton, Colorado, USA). .. Extracted proteins were visualized using Amersham ECL Prime Western Blotting Detection Reagent (#RPN2232, GE Healthcare Life Sciences, Little Chalfont, United Kingdom).

Article Title: Increased plasma VEGF levels following ischemic preconditioning are associated with downregulation of miRNA-762 and miR-3072-5p
Article Snippet: Cells were re-suspended in cold buffer containing 10-mM Tris-HCl (pH 7.4), 0.5-mM EGTA, 0.5-mM EDTA, 1% Triton X-100, and cOmpleteTM Mini Protease Inhibitor Cocktail (Sigma, St Louis, MO, USA) and were incubated for 30 min on ice and then centrifuged at 12 000 g for 20 min at 4 °C. .. Supernatants containing total protein were collected and subjected to Western blotting using rabbit anti-GDF15/MIC-polyclonal antibody (1:500 dilution, # bs-3818R, Bioss, Woburn, Massachusetts, USA), anti-rabbit IgG HRP-conjugated secondary antibody (1:5000 dilution, #P0448, Dako, Glostrup, Denmark), and anti-β-actin HRP-conjugated antibody (1:2000 dilution #NB600-532H, Novus Biologicals, Littleton, Colorado, USA). .. Extracted proteins were visualized using Amersham ECL Prime Western Blotting Detection Reagent (#RPN2232, GE Healthcare Life Sciences, Little Chalfont, United Kingdom).

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Correlations between GDF15 IHC score and clinical characteristics in patients with breast cancer

Journal: Oncotarget

Article Title: Growth differentiation factor 15 mediates epithelial mesenchymal transition and invasion of breast cancers through IGF-1R-FoxM1 signaling

doi: 10.18632/oncotarget.21765

Figure Lengend Snippet: Correlations between GDF15 IHC score and clinical characteristics in patients with breast cancer

Article Snippet: TMAs were then stained with rabbit polyclonal anti-human GDF15 (also known as MIC-1; antibody 3249; dilution 1:100; Cell Signaling Technology, Inc., 3 Trask Lane, Danvers, MA 01923) overnight at 4°C.

Techniques:

(A) Western blotting ( above ) and real-time PCR ( graph ) for GDF15 in BT474, JIMT1 and MDA-MB-231 (MDA231) breast cancer cell lines. PCR values reflect fold change in GDF15 transcript normalized to RPLPO housekeeping gene. Error bars represent standard deviation between triplicate samples; experiments were repeated at least 3 times. (B) Real-time PCR for GDF15 in BT474 pCMV stable empty vector control clone (pCMV) and GDF15 stable clones 2, 3, and 5 (C2, C3, and C5). Values reflect fold change in GDF15 transcript level normalized to RPLPO housekeeping gene. Error bars represent standard deviation between triplicate samples; experiments were repeated at least 3 times. (C-D) BT474 parental, pCMV empty vector control clone (pCMV), and GDF15 stable clones 2 and 3 (C2, C3) were fixed, stained with propidium iodide, and analyzed for DNA content by flow cytometry. The percentage of cells in each cell cycle phase is shown per cell line (C) (white, G0/G1; gray, S; black, G2/M). Error bars represent standard deviation between triplicate samples; experiments were repeated at least 3 times. Representative cell cycle histograms are shown per line (D).

Journal: Oncotarget

Article Title: Growth differentiation factor 15 mediates epithelial mesenchymal transition and invasion of breast cancers through IGF-1R-FoxM1 signaling

doi: 10.18632/oncotarget.21765

Figure Lengend Snippet: (A) Western blotting ( above ) and real-time PCR ( graph ) for GDF15 in BT474, JIMT1 and MDA-MB-231 (MDA231) breast cancer cell lines. PCR values reflect fold change in GDF15 transcript normalized to RPLPO housekeeping gene. Error bars represent standard deviation between triplicate samples; experiments were repeated at least 3 times. (B) Real-time PCR for GDF15 in BT474 pCMV stable empty vector control clone (pCMV) and GDF15 stable clones 2, 3, and 5 (C2, C3, and C5). Values reflect fold change in GDF15 transcript level normalized to RPLPO housekeeping gene. Error bars represent standard deviation between triplicate samples; experiments were repeated at least 3 times. (C-D) BT474 parental, pCMV empty vector control clone (pCMV), and GDF15 stable clones 2 and 3 (C2, C3) were fixed, stained with propidium iodide, and analyzed for DNA content by flow cytometry. The percentage of cells in each cell cycle phase is shown per cell line (C) (white, G0/G1; gray, S; black, G2/M). Error bars represent standard deviation between triplicate samples; experiments were repeated at least 3 times. Representative cell cycle histograms are shown per line (D).

Techniques: Western Blot, Real-time Polymerase Chain Reaction, Standard Deviation, Plasmid Preparation, Control, Clone Assay, Staining, Flow Cytometry

(A) Total protein whole-cell lysates were collected from BT474 pCMV stable empty vector control clone (pCMV) and GDF15 stable clones 2, 3, and 5 (C2, C3, and C5). Western blots were performed for E-Cadherin, N-Cadherin, vimentin, and FoxM1; actin was measured as a loading control. Blots were repeated at least three times, and representative blots are shown. (B) Real-time PCR for Snail, Zeb-1 and Slug in BT474 parental, pCMV stable empty vector control clone (pCMV) and GDF15 stable clones 2, 3, and 5 (C2, C3, and C5). Values reflect fold change in transcript normalized to RPLPO housekeeping gene. Error bars represent standard deviation between triplicate samples; experiments were repeated at least 3 times. (C) BT474 pCMV empty vector control clone (pCMV) and GDF15 stable clones 3 and 5 (C3 and C5) were imaged at 10× magnification to evaluate changes in morphology. (D) Representative images of spheroid cultures are shown for BT474 parental and GDF15 stable clone 2 (C2). (E) BT474 stable empty vector control clone (pCMV) and GDF15 stable clones 2, 3, and 5 (C2, C3, and C5) were plated in basement membrane matrix mimic (Matrigel)-coated Boyden chambers in serum-free media; 10% FBS was added to the well below each chamber as a chemo-attractant. After 24 hours, cells were fixed and stained. Representative photos of invading cells are shown at 20× magnification. The total number of invading cells was counted in 10 random fields; the average number of invading cells is shown for triplicate cultures per cell line; student’s t-test, ** p<0.005, * p<0.05.

Journal: Oncotarget

Article Title: Growth differentiation factor 15 mediates epithelial mesenchymal transition and invasion of breast cancers through IGF-1R-FoxM1 signaling

doi: 10.18632/oncotarget.21765

Figure Lengend Snippet: (A) Total protein whole-cell lysates were collected from BT474 pCMV stable empty vector control clone (pCMV) and GDF15 stable clones 2, 3, and 5 (C2, C3, and C5). Western blots were performed for E-Cadherin, N-Cadherin, vimentin, and FoxM1; actin was measured as a loading control. Blots were repeated at least three times, and representative blots are shown. (B) Real-time PCR for Snail, Zeb-1 and Slug in BT474 parental, pCMV stable empty vector control clone (pCMV) and GDF15 stable clones 2, 3, and 5 (C2, C3, and C5). Values reflect fold change in transcript normalized to RPLPO housekeeping gene. Error bars represent standard deviation between triplicate samples; experiments were repeated at least 3 times. (C) BT474 pCMV empty vector control clone (pCMV) and GDF15 stable clones 3 and 5 (C3 and C5) were imaged at 10× magnification to evaluate changes in morphology. (D) Representative images of spheroid cultures are shown for BT474 parental and GDF15 stable clone 2 (C2). (E) BT474 stable empty vector control clone (pCMV) and GDF15 stable clones 2, 3, and 5 (C2, C3, and C5) were plated in basement membrane matrix mimic (Matrigel)-coated Boyden chambers in serum-free media; 10% FBS was added to the well below each chamber as a chemo-attractant. After 24 hours, cells were fixed and stained. Representative photos of invading cells are shown at 20× magnification. The total number of invading cells was counted in 10 random fields; the average number of invading cells is shown for triplicate cultures per cell line; student’s t-test, ** p<0.005, * p<0.05.

Techniques: Plasmid Preparation, Control, Clone Assay, Western Blot, Real-time Polymerase Chain Reaction, Standard Deviation, Stable Transfection, Membrane, Staining

(A) Total protein lysates were collected from BT474 stable empty vector control clone (pCMV) and GDF15 stable clones 2, 3, and 5 (C2, C3, and C5). Western blots were performed for p-Tyr1131 IGF-1R and total IGF-1R; actin was probed as loading control. Experiments were repeated 3 times; representative blots are shown. Quantification (shown beneath each band) was normalized to actin and performed using Odyssey Li-Cor imaging software. (B) BT474 GDF15 stable clone 2 (C2) cells were treated with normal mouse IgG control or 0.25 μg/mL alpha IR3 (aIR3) IGF-1R monoclonal antibody for 48 hours. Western blots of total protein lysates were performed for total IGF-1R, vimentin, FoxM1, and E-Cadherin. Bar graphs show quantification relative to actin loading control, and was performed using Odyssey Li-Cor imaging software. Error bars represent standard deviation between triplicates; experiments were performed at least 3 times. (C) BT474 stable empty vector control clone (pCMV) and BT474 GDF15 stable clone 5 (C5) cells were pre-treated with normal mouse IgG or 0.25 μg/mL alpha IR3 (aIR3) IGF-1R monoclonal antibody for 24 hours. Cells were then seeded in Matrigel-coated Boyden chambers in serum-free media plus control IgG or aIR3; 10% FBS was added to the well as a chemo-attractant. After 24 hours of invasion, cells in chambers were fixed and stained. Representative photos of invading cells are shown at 20× magnification. The total number of invading cells was counted in 10 random fields; the average number of invading cells is shown for triplicate cultures per cell line, * p<0.05.

Journal: Oncotarget

Article Title: Growth differentiation factor 15 mediates epithelial mesenchymal transition and invasion of breast cancers through IGF-1R-FoxM1 signaling

doi: 10.18632/oncotarget.21765

Figure Lengend Snippet: (A) Total protein lysates were collected from BT474 stable empty vector control clone (pCMV) and GDF15 stable clones 2, 3, and 5 (C2, C3, and C5). Western blots were performed for p-Tyr1131 IGF-1R and total IGF-1R; actin was probed as loading control. Experiments were repeated 3 times; representative blots are shown. Quantification (shown beneath each band) was normalized to actin and performed using Odyssey Li-Cor imaging software. (B) BT474 GDF15 stable clone 2 (C2) cells were treated with normal mouse IgG control or 0.25 μg/mL alpha IR3 (aIR3) IGF-1R monoclonal antibody for 48 hours. Western blots of total protein lysates were performed for total IGF-1R, vimentin, FoxM1, and E-Cadherin. Bar graphs show quantification relative to actin loading control, and was performed using Odyssey Li-Cor imaging software. Error bars represent standard deviation between triplicates; experiments were performed at least 3 times. (C) BT474 stable empty vector control clone (pCMV) and BT474 GDF15 stable clone 5 (C5) cells were pre-treated with normal mouse IgG or 0.25 μg/mL alpha IR3 (aIR3) IGF-1R monoclonal antibody for 24 hours. Cells were then seeded in Matrigel-coated Boyden chambers in serum-free media plus control IgG or aIR3; 10% FBS was added to the well as a chemo-attractant. After 24 hours of invasion, cells in chambers were fixed and stained. Representative photos of invading cells are shown at 20× magnification. The total number of invading cells was counted in 10 random fields; the average number of invading cells is shown for triplicate cultures per cell line, * p<0.05.

Techniques: Plasmid Preparation, Control, Clone Assay, Western Blot, Imaging, Software, Stable Transfection, Standard Deviation, Staining

(A) Real-time PCR of total RNA from BT474 pCMV empty vector control clone (pCMV) and GDF15 stable clones (C2, C3, and C5) for MMP2 ( left graph ) and MMP9 ( right graph ). Values reflect average fold in transcript normalized to internal control RPLPO relative to pCMV group. Error bars represent standard deviation between triplicate samples; experiments were repeated 3 times. (B) BT474 pCMV empty vector control clone (pCMV) and GDF15 stable clones (C2, C3 and C5) were plated in serum-free media in Matrigel-coated Boyden chambers and treated with vehicle control or 1 μg pan-MMP inhibitor GM6001 for 24 hours, after which cells were fixed and stained. Representative photos of invading cells are shown at 20× magnification. The total number of invading cells was counted in 10 random fields; the average number of invading cells is shown for triplicate cultures per cell line; student’s t-test, ** p<0.005, * p<0.05.

Journal: Oncotarget

Article Title: Growth differentiation factor 15 mediates epithelial mesenchymal transition and invasion of breast cancers through IGF-1R-FoxM1 signaling

doi: 10.18632/oncotarget.21765

Figure Lengend Snippet: (A) Real-time PCR of total RNA from BT474 pCMV empty vector control clone (pCMV) and GDF15 stable clones (C2, C3, and C5) for MMP2 ( left graph ) and MMP9 ( right graph ). Values reflect average fold in transcript normalized to internal control RPLPO relative to pCMV group. Error bars represent standard deviation between triplicate samples; experiments were repeated 3 times. (B) BT474 pCMV empty vector control clone (pCMV) and GDF15 stable clones (C2, C3 and C5) were plated in serum-free media in Matrigel-coated Boyden chambers and treated with vehicle control or 1 μg pan-MMP inhibitor GM6001 for 24 hours, after which cells were fixed and stained. Representative photos of invading cells are shown at 20× magnification. The total number of invading cells was counted in 10 random fields; the average number of invading cells is shown for triplicate cultures per cell line; student’s t-test, ** p<0.005, * p<0.05.

Techniques: Real-time Polymerase Chain Reaction, Plasmid Preparation, Control, Clone Assay, Standard Deviation, Staining

(A) BT474 stable empty vector control clone (pCMV) and GDF15 stable clone 5 (C5) were transfected with 100 nM control siRNA (siCtrl) or FoxM1 siRNA (siFoxM1) for 48 hours, and then plated in serum-free media in Matrigel-coated Boyden chambers. After 24 hours, cells were fixed and stained. Representative photos of invading cells are shown at 20× magnification. The total number of invading cells was counted in 10 random fields; the average number of invading cells is shown for triplicate cultures per cell line; student’s t-test, ** p<0.005. (B) BT474 GDF15 stable clone 5 (C5) cells were transfected with 100 nM control siRNA (siCtrl) or FoxM1 siRNA (FoxM1) for 48 hours. Real-time PCR was performed for FoxM1, MMP2, and MMP9. Values reflect average fold in transcript normalized to internal control RPLPO. Error bars represent standard deviation between triplicate samples; experiments were repeated 3 times, ** p<0.005.

Journal: Oncotarget

Article Title: Growth differentiation factor 15 mediates epithelial mesenchymal transition and invasion of breast cancers through IGF-1R-FoxM1 signaling

doi: 10.18632/oncotarget.21765

Figure Lengend Snippet: (A) BT474 stable empty vector control clone (pCMV) and GDF15 stable clone 5 (C5) were transfected with 100 nM control siRNA (siCtrl) or FoxM1 siRNA (siFoxM1) for 48 hours, and then plated in serum-free media in Matrigel-coated Boyden chambers. After 24 hours, cells were fixed and stained. Representative photos of invading cells are shown at 20× magnification. The total number of invading cells was counted in 10 random fields; the average number of invading cells is shown for triplicate cultures per cell line; student’s t-test, ** p<0.005. (B) BT474 GDF15 stable clone 5 (C5) cells were transfected with 100 nM control siRNA (siCtrl) or FoxM1 siRNA (FoxM1) for 48 hours. Real-time PCR was performed for FoxM1, MMP2, and MMP9. Values reflect average fold in transcript normalized to internal control RPLPO. Error bars represent standard deviation between triplicate samples; experiments were repeated 3 times, ** p<0.005.

Techniques: Plasmid Preparation, Control, Stable Transfection, Transfection, Staining, Real-time Polymerase Chain Reaction, Standard Deviation

(A) JIMT1 cells were serum starved for 24 hours, and then stimulated with 2 or 20 ng/mL of recombinant human GDF15 (rhGDF15) for another 24 hours. Western blots of total protein lysates are shown for p-Tyr1131 IGF-1R and total IGF-1R; actin was probed as loading control. Experiments were repeated 3 times; representative blots are shown. Quantification (shown beneath each band) was normalized to actin and performed using Odyssey Li-Cor imaging software. (B) JIMT-1 cells were pretreated for 24 hours with control IgG or 0.25 μg/ml alpha IR3 (aIR3) in serum-free media, and then stimulated with 2 or 20 ng/mL of recombinant human GDF15 (rhGDF15) for another 24 hours. Western blots of total protein lysates are shown for total IGF-1R with actin as loading control. Experiments were repeated 3 times; representative blots are shown. Quantification (shown beneath each band) was normalized to actin and performed using Odyssey Li-Cor imaging software. (C) JIMT-1 cells were pretreated for 24 hours with control IgG or 0.25 μg/ml alpha IR3 (aIR3) in serum-free media, and then stimulated with 2 or 20 ng/mL of recombinant human GDF15 (rhGDF15) for another 24 hours. Real-time PCR was performed for SNAIL, SLUG, MMP2, and MMP9. Values reflect average fold in transcript normalized to internal control RPLPO. Error bars represent standard deviation between triplicate samples; experiments were repeated 3 times. (D) JIMT-1 cells were pretreated for 24 hours with control IgG or 0.25 μg/ml alpha IR3 (aIR3) in serum-free media, and then seeded in Matrigel-coated Boyden chambers in serum-free media with 10% FBS in the lower chamber as chemoattractant. Drug treatment was continued (in indicated samples), and 2 or 20 ng/mL of recombinant human GDF15 (rhGDF15) was added to the chambers of treatment groups where indicated. After 24 hours of invasion, photos were taken at 20× magnification; representative photos are shown. The number of invaded cells is shown per group; error bars represent standard deviation between triplicate samples, * p<0.05.

Journal: Oncotarget

Article Title: Growth differentiation factor 15 mediates epithelial mesenchymal transition and invasion of breast cancers through IGF-1R-FoxM1 signaling

doi: 10.18632/oncotarget.21765

Figure Lengend Snippet: (A) JIMT1 cells were serum starved for 24 hours, and then stimulated with 2 or 20 ng/mL of recombinant human GDF15 (rhGDF15) for another 24 hours. Western blots of total protein lysates are shown for p-Tyr1131 IGF-1R and total IGF-1R; actin was probed as loading control. Experiments were repeated 3 times; representative blots are shown. Quantification (shown beneath each band) was normalized to actin and performed using Odyssey Li-Cor imaging software. (B) JIMT-1 cells were pretreated for 24 hours with control IgG or 0.25 μg/ml alpha IR3 (aIR3) in serum-free media, and then stimulated with 2 or 20 ng/mL of recombinant human GDF15 (rhGDF15) for another 24 hours. Western blots of total protein lysates are shown for total IGF-1R with actin as loading control. Experiments were repeated 3 times; representative blots are shown. Quantification (shown beneath each band) was normalized to actin and performed using Odyssey Li-Cor imaging software. (C) JIMT-1 cells were pretreated for 24 hours with control IgG or 0.25 μg/ml alpha IR3 (aIR3) in serum-free media, and then stimulated with 2 or 20 ng/mL of recombinant human GDF15 (rhGDF15) for another 24 hours. Real-time PCR was performed for SNAIL, SLUG, MMP2, and MMP9. Values reflect average fold in transcript normalized to internal control RPLPO. Error bars represent standard deviation between triplicate samples; experiments were repeated 3 times. (D) JIMT-1 cells were pretreated for 24 hours with control IgG or 0.25 μg/ml alpha IR3 (aIR3) in serum-free media, and then seeded in Matrigel-coated Boyden chambers in serum-free media with 10% FBS in the lower chamber as chemoattractant. Drug treatment was continued (in indicated samples), and 2 or 20 ng/mL of recombinant human GDF15 (rhGDF15) was added to the chambers of treatment groups where indicated. After 24 hours of invasion, photos were taken at 20× magnification; representative photos are shown. The number of invaded cells is shown per group; error bars represent standard deviation between triplicate samples, * p<0.05.

Techniques: Recombinant, Western Blot, Control, Imaging, Software, Real-time Polymerase Chain Reaction, Standard Deviation

(A) JIMT-1 cells were transfected with 100 nM control siRNA (siCtrl) or FoxM1 siRNA (siFoxM1) for 24 hours, and then stimulated with 2 or 20 ng/mL recombinant human GDF15 (rhGDF15) for another 24 hours. Western blots of total protein lysates are shown for total FoxM1 with actin as loading control. (B) JIMT-1 cells were transfected with 100 nM control siRNA (siCtrl) or FoxM1 siRNA (siFoxM1) for 24 hours, and then stimulated with 2 or 20 ng/mL recombinant human GDF15 (rhGDF15) for another 24 hours. Real-time PCR was performed for Snail, Slug, MMP2, and MMP9. Values reflect average fold in transcript normalized to internal control RPLPO. Error bars represent standard deviation between triplicate samples; experiments were repeated 3 times. (C) JIMT-1 cells were transfected with 100 nM control siRNA (siCtrl) or FoxM1 siRNA (siFoxM1) for 24 hours, and then plated in serum-free media in Matrigel-coated Boyden chambers. Recombinant human GDF15 (rhGDF15; 2 or 20 ng/mL) was added to lower chambers where indicated. After 24 hours, cells were fixed and stained; representative photos of invading cells are shown at 20× magnification. The total number of invading cells was counted in 10 random fields; the average number of invading cells is shown for triplicate cultures per cell line, ** p<0.005, * p<0.05.

Journal: Oncotarget

Article Title: Growth differentiation factor 15 mediates epithelial mesenchymal transition and invasion of breast cancers through IGF-1R-FoxM1 signaling

doi: 10.18632/oncotarget.21765

Figure Lengend Snippet: (A) JIMT-1 cells were transfected with 100 nM control siRNA (siCtrl) or FoxM1 siRNA (siFoxM1) for 24 hours, and then stimulated with 2 or 20 ng/mL recombinant human GDF15 (rhGDF15) for another 24 hours. Western blots of total protein lysates are shown for total FoxM1 with actin as loading control. (B) JIMT-1 cells were transfected with 100 nM control siRNA (siCtrl) or FoxM1 siRNA (siFoxM1) for 24 hours, and then stimulated with 2 or 20 ng/mL recombinant human GDF15 (rhGDF15) for another 24 hours. Real-time PCR was performed for Snail, Slug, MMP2, and MMP9. Values reflect average fold in transcript normalized to internal control RPLPO. Error bars represent standard deviation between triplicate samples; experiments were repeated 3 times. (C) JIMT-1 cells were transfected with 100 nM control siRNA (siCtrl) or FoxM1 siRNA (siFoxM1) for 24 hours, and then plated in serum-free media in Matrigel-coated Boyden chambers. Recombinant human GDF15 (rhGDF15; 2 or 20 ng/mL) was added to lower chambers where indicated. After 24 hours, cells were fixed and stained; representative photos of invading cells are shown at 20× magnification. The total number of invading cells was counted in 10 random fields; the average number of invading cells is shown for triplicate cultures per cell line, ** p<0.005, * p<0.05.

Techniques: Transfection, Control, Recombinant, Western Blot, Real-time Polymerase Chain Reaction, Standard Deviation, Staining

(A) BT474 stable empty vector control clone (pCMV), BT474 GDF15 stable clone 5 (C5), and MDA-MB-231 cells were transfected with 100 nM siRNA control (siCtrl) or GDF15 siRNA (siGDF15). After 24 hours, transfected cells were plated in serum-free media in Matrigel-coated Boyden chambers with 10% FBS in the wells as a chemoattractant. After 24 hours, cells were fixed and stained. Representative photos of invading cells are shown at 20× magnification. The total number of invading cells was counted in 10 random fields; the average number of invading cells is shown for triplicate cultures per cell line, * p<0.05. (B) BT474 stable empty vector control clone (pCMV), BT474 GDF15 stable clone 5 (C5), and MDA-MB-231 cells were transfected with 100 nM siRNA control (siCtrl) or GDF15 siRNA (siGDF15) for 48 hours. Real-time PCR was performed to confirm GDF15 knockdown. Values reflect the fold change in transcript normalized to RPLPO housekeeping gene. Error bars represent standard deviation between triplicate samples; experiments were repeated twice; ** p<0.005, * p<0.05.

Journal: Oncotarget

Article Title: Growth differentiation factor 15 mediates epithelial mesenchymal transition and invasion of breast cancers through IGF-1R-FoxM1 signaling

doi: 10.18632/oncotarget.21765

Figure Lengend Snippet: (A) BT474 stable empty vector control clone (pCMV), BT474 GDF15 stable clone 5 (C5), and MDA-MB-231 cells were transfected with 100 nM siRNA control (siCtrl) or GDF15 siRNA (siGDF15). After 24 hours, transfected cells were plated in serum-free media in Matrigel-coated Boyden chambers with 10% FBS in the wells as a chemoattractant. After 24 hours, cells were fixed and stained. Representative photos of invading cells are shown at 20× magnification. The total number of invading cells was counted in 10 random fields; the average number of invading cells is shown for triplicate cultures per cell line, * p<0.05. (B) BT474 stable empty vector control clone (pCMV), BT474 GDF15 stable clone 5 (C5), and MDA-MB-231 cells were transfected with 100 nM siRNA control (siCtrl) or GDF15 siRNA (siGDF15) for 48 hours. Real-time PCR was performed to confirm GDF15 knockdown. Values reflect the fold change in transcript normalized to RPLPO housekeeping gene. Error bars represent standard deviation between triplicate samples; experiments were repeated twice; ** p<0.005, * p<0.05.

Techniques: Plasmid Preparation, Control, Stable Transfection, Transfection, Staining, Real-time Polymerase Chain Reaction, Knockdown, Standard Deviation

GDF15 activates IGF-1R signaling and induces expression of FoxM1, which upregulates expression of EMT transcription factors (TFs), including Snail and Slug, and induces expression of matrix metalloproteinases (MMPs), stimulating EMT and breast cancer cell invasion.

Journal: Oncotarget

Article Title: Growth differentiation factor 15 mediates epithelial mesenchymal transition and invasion of breast cancers through IGF-1R-FoxM1 signaling

doi: 10.18632/oncotarget.21765

Figure Lengend Snippet: GDF15 activates IGF-1R signaling and induces expression of FoxM1, which upregulates expression of EMT transcription factors (TFs), including Snail and Slug, and induces expression of matrix metalloproteinases (MMPs), stimulating EMT and breast cancer cell invasion.

Techniques: Expressing

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