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Ortho-PDX generation is associated with TGF-β pathway activation and increased expression of stemness-associated factors in the parental tumor. (A) Left: immunoblot analysis of TGF-β, LTBP-1 (Latent-transforming growth factor beta-binding protein 1), Latent TGF-β, active TGF-β and phosphorylated SMAD <t>2/3</t> <t>(pSMAD</t> 2/3) on whole lysates of normal mucosae (N) and patient tumors (T). Tubulin and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were used as loading controls. Right: quantification of the immunoblot experiment shown on the left. (B) ELISA assay performed on lysates of normal mucosae (N) and patient tumors (T) showing active TGF-β concentration. ***P < 0.001 from two-tailed t test. (C) pSMAD 2/3 staining of L5, L6 and L7 tumor sections and quantification of 5 fields/section. Magnification 20x. Bar 50 μm. (D) qRT-PCR analysis of Bmi-1, Nanog and LGR5 expression in normal mucosae and tumor tissues of CRC patients. Mean ± SD of 3 experiments. *P < 0.05, **P < 0.01 and ***P < 0.001 from two-tailed t test.
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Ortho-PDX generation is associated with TGF-β pathway activation and increased expression of stemness-associated factors in the parental tumor. (A) Left: immunoblot analysis of TGF-β, LTBP-1 (Latent-transforming growth factor beta-binding protein 1), Latent TGF-β, active TGF-β and phosphorylated SMAD <t>2/3</t> <t>(pSMAD</t> 2/3) on whole lysates of normal mucosae (N) and patient tumors (T). Tubulin and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were used as loading controls. Right: quantification of the immunoblot experiment shown on the left. (B) ELISA assay performed on lysates of normal mucosae (N) and patient tumors (T) showing active TGF-β concentration. ***P < 0.001 from two-tailed t test. (C) pSMAD 2/3 staining of L5, L6 and L7 tumor sections and quantification of 5 fields/section. Magnification 20x. Bar 50 μm. (D) qRT-PCR analysis of Bmi-1, Nanog and LGR5 expression in normal mucosae and tumor tissues of CRC patients. Mean ± SD of 3 experiments. *P < 0.05, **P < 0.01 and ***P < 0.001 from two-tailed t test.
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Ortho-PDX generation is associated with TGF-β pathway activation and increased expression of stemness-associated factors in the parental tumor. (A) Left: immunoblot analysis of TGF-β, LTBP-1 (Latent-transforming growth factor beta-binding protein 1), Latent TGF-β, active TGF-β and phosphorylated SMAD <t>2/3</t> <t>(pSMAD</t> 2/3) on whole lysates of normal mucosae (N) and patient tumors (T). Tubulin and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were used as loading controls. Right: quantification of the immunoblot experiment shown on the left. (B) ELISA assay performed on lysates of normal mucosae (N) and patient tumors (T) showing active TGF-β concentration. ***P < 0.001 from two-tailed t test. (C) pSMAD 2/3 staining of L5, L6 and L7 tumor sections and quantification of 5 fields/section. Magnification 20x. Bar 50 μm. (D) qRT-PCR analysis of Bmi-1, Nanog and LGR5 expression in normal mucosae and tumor tissues of CRC patients. Mean ± SD of 3 experiments. *P < 0.05, **P < 0.01 and ***P < 0.001 from two-tailed t test.
Square Well Corning 3711 Microtiter Plates, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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CodonCode corporation aligner 3.7.1.1 sequencing analysis software
Ortho-PDX generation is associated with TGF-β pathway activation and increased expression of stemness-associated factors in the parental tumor. (A) Left: immunoblot analysis of TGF-β, LTBP-1 (Latent-transforming growth factor beta-binding protein 1), Latent TGF-β, active TGF-β and phosphorylated SMAD <t>2/3</t> <t>(pSMAD</t> 2/3) on whole lysates of normal mucosae (N) and patient tumors (T). Tubulin and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were used as loading controls. Right: quantification of the immunoblot experiment shown on the left. (B) ELISA assay performed on lysates of normal mucosae (N) and patient tumors (T) showing active TGF-β concentration. ***P < 0.001 from two-tailed t test. (C) pSMAD 2/3 staining of L5, L6 and L7 tumor sections and quantification of 5 fields/section. Magnification 20x. Bar 50 μm. (D) qRT-PCR analysis of Bmi-1, Nanog and LGR5 expression in normal mucosae and tumor tissues of CRC patients. Mean ± SD of 3 experiments. *P < 0.05, **P < 0.01 and ***P < 0.001 from two-tailed t test.
Aligner 3.7.1.1 Sequencing Analysis Software, supplied by CodonCode corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Ortho-PDX generation is associated with TGF-β pathway activation and increased expression of stemness-associated factors in the parental tumor. (A) Left: immunoblot analysis of TGF-β, LTBP-1 (Latent-transforming growth factor beta-binding protein 1), Latent TGF-β, active TGF-β and phosphorylated SMAD <t>2/3</t> <t>(pSMAD</t> 2/3) on whole lysates of normal mucosae (N) and patient tumors (T). Tubulin and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were used as loading controls. Right: quantification of the immunoblot experiment shown on the left. (B) ELISA assay performed on lysates of normal mucosae (N) and patient tumors (T) showing active TGF-β concentration. ***P < 0.001 from two-tailed t test. (C) pSMAD 2/3 staining of L5, L6 and L7 tumor sections and quantification of 5 fields/section. Magnification 20x. Bar 50 μm. (D) qRT-PCR analysis of Bmi-1, Nanog and LGR5 expression in normal mucosae and tumor tissues of CRC patients. Mean ± SD of 3 experiments. *P < 0.05, **P < 0.01 and ***P < 0.001 from two-tailed t test.
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Image Search Results


Ortho-PDX generation is associated with TGF-β pathway activation and increased expression of stemness-associated factors in the parental tumor. (A) Left: immunoblot analysis of TGF-β, LTBP-1 (Latent-transforming growth factor beta-binding protein 1), Latent TGF-β, active TGF-β and phosphorylated SMAD 2/3 (pSMAD 2/3) on whole lysates of normal mucosae (N) and patient tumors (T). Tubulin and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were used as loading controls. Right: quantification of the immunoblot experiment shown on the left. (B) ELISA assay performed on lysates of normal mucosae (N) and patient tumors (T) showing active TGF-β concentration. ***P < 0.001 from two-tailed t test. (C) pSMAD 2/3 staining of L5, L6 and L7 tumor sections and quantification of 5 fields/section. Magnification 20x. Bar 50 μm. (D) qRT-PCR analysis of Bmi-1, Nanog and LGR5 expression in normal mucosae and tumor tissues of CRC patients. Mean ± SD of 3 experiments. *P < 0.05, **P < 0.01 and ***P < 0.001 from two-tailed t test.

Journal: Frontiers in Oncology

Article Title: An Orthotopic Patient-Derived Xenograft (PDX) Model Allows the Analysis of Metastasis-Associated Features in Colorectal Cancer

doi: 10.3389/fonc.2022.869485

Figure Lengend Snippet: Ortho-PDX generation is associated with TGF-β pathway activation and increased expression of stemness-associated factors in the parental tumor. (A) Left: immunoblot analysis of TGF-β, LTBP-1 (Latent-transforming growth factor beta-binding protein 1), Latent TGF-β, active TGF-β and phosphorylated SMAD 2/3 (pSMAD 2/3) on whole lysates of normal mucosae (N) and patient tumors (T). Tubulin and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were used as loading controls. Right: quantification of the immunoblot experiment shown on the left. (B) ELISA assay performed on lysates of normal mucosae (N) and patient tumors (T) showing active TGF-β concentration. ***P < 0.001 from two-tailed t test. (C) pSMAD 2/3 staining of L5, L6 and L7 tumor sections and quantification of 5 fields/section. Magnification 20x. Bar 50 μm. (D) qRT-PCR analysis of Bmi-1, Nanog and LGR5 expression in normal mucosae and tumor tissues of CRC patients. Mean ± SD of 3 experiments. *P < 0.05, **P < 0.01 and ***P < 0.001 from two-tailed t test.

Article Snippet: Vimentin (#5741), N-Cadherin (#13116), SLUG (#9585), ZEB1 (#3396), pSMAD 2/3 (#8828), TGFβ (#3711) were from Cell Signaling Technology, E-Cadherin (#610181) from Becton Dickinson, LTBP1 from Santa Cruz (#sc-271140).

Techniques: Activation Assay, Expressing, Western Blot, Binding Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, Two Tailed Test, Staining, Quantitative RT-PCR

Metastatic tissues lose mesenchymal traits and acquire increased chemoresistance. (A) Left: comparative immunoblot analysis of E-Cadherin (E-CAD) and Vimentin (VIM) on whole lysates of normal/peritumoral mucosa (N) and tumor tissue (T) of patient L6, orthotopic xenograft (Orthotopic), liver and lung xenograft metastases (Liver Met and Lung Met). GAPDH was used as a loading control. Right: quantification of the immunoblot experiment shown on the left. (B) Left: comparative immunoblot analysis of phosphorylated SMAD 2/3 (pSMAD 2/3) and TGF-β on whole lysates as described above. Right: quantification of the immunoblot experiment shown on the left. (C) qRT-PCR analysis of E-Cadherin (E-CAD), Vimentin (VIM), Twist, ZEB1, SNAIL, SLUG, and ZEB2 in normal/peritumoral mucosa (N) and tumor tissue (T) of patient L6, orthotopic xenograft (Orthotopic), liver and lung xenograft metastases (Liver Met and Lung Met). Mean ± SD of 3 experiments. *P < 0.05, **P < 0.01 and ***P < 0.001 from two-tailed Student’s t test. (D) Staining of alpha Smooth Muscle Actin (αSMA) on sections of L6 orthotopic PDX and hepatic metastasis (left) and quantification of 5 sections (right). Magnification 20x, ns non-significant. (E) Left: Immunoblot analysis of human Fibroblast Activation Protein (FAP) in normal (N) and tumor (T) parental tissues of L6 patient, in orthotopic PDX (Orthotopic) and metastatic PDX (Liver Met and Lung Met). Tubulin was used as loading control. Right: quantification of the immunoblot experiment shown on the left. (F) Time course of organoid generation (orthotopic xenograft-derived organoids, OXDOs and metastatic xenograft-derived organoids, MXDOs), from the first day of culture (passage 0, P0) to subsequent passages (P3, P5 and P7, respectively after 3 weeks, 5 weeks and 7 weeks of culture). Magnification 10x. Bar 100 μM. (G) qRT-PCR analysis of E-Cadherin (E-CAD), Vimentin (VIM), SNAIL and ZEB2 on OXDOs (orange bars) and MXDOs (blue bars). *P < 0.05, **P < 0.01 and ***P < 0.001 from two-tailed Student’s t test. (H) Invasion/migration assay performed with OXDOs and MXDOs. (I) Cell viability of OXDOs (orange bars) and MXDOs (blue bars) treated with 5 μM 5-Fluorouracil (5-FU) for 6 days. Values represent mean ± SD of three independent experiments. *P < 0.05 by unpaired Student’s t test.

Journal: Frontiers in Oncology

Article Title: An Orthotopic Patient-Derived Xenograft (PDX) Model Allows the Analysis of Metastasis-Associated Features in Colorectal Cancer

doi: 10.3389/fonc.2022.869485

Figure Lengend Snippet: Metastatic tissues lose mesenchymal traits and acquire increased chemoresistance. (A) Left: comparative immunoblot analysis of E-Cadherin (E-CAD) and Vimentin (VIM) on whole lysates of normal/peritumoral mucosa (N) and tumor tissue (T) of patient L6, orthotopic xenograft (Orthotopic), liver and lung xenograft metastases (Liver Met and Lung Met). GAPDH was used as a loading control. Right: quantification of the immunoblot experiment shown on the left. (B) Left: comparative immunoblot analysis of phosphorylated SMAD 2/3 (pSMAD 2/3) and TGF-β on whole lysates as described above. Right: quantification of the immunoblot experiment shown on the left. (C) qRT-PCR analysis of E-Cadherin (E-CAD), Vimentin (VIM), Twist, ZEB1, SNAIL, SLUG, and ZEB2 in normal/peritumoral mucosa (N) and tumor tissue (T) of patient L6, orthotopic xenograft (Orthotopic), liver and lung xenograft metastases (Liver Met and Lung Met). Mean ± SD of 3 experiments. *P < 0.05, **P < 0.01 and ***P < 0.001 from two-tailed Student’s t test. (D) Staining of alpha Smooth Muscle Actin (αSMA) on sections of L6 orthotopic PDX and hepatic metastasis (left) and quantification of 5 sections (right). Magnification 20x, ns non-significant. (E) Left: Immunoblot analysis of human Fibroblast Activation Protein (FAP) in normal (N) and tumor (T) parental tissues of L6 patient, in orthotopic PDX (Orthotopic) and metastatic PDX (Liver Met and Lung Met). Tubulin was used as loading control. Right: quantification of the immunoblot experiment shown on the left. (F) Time course of organoid generation (orthotopic xenograft-derived organoids, OXDOs and metastatic xenograft-derived organoids, MXDOs), from the first day of culture (passage 0, P0) to subsequent passages (P3, P5 and P7, respectively after 3 weeks, 5 weeks and 7 weeks of culture). Magnification 10x. Bar 100 μM. (G) qRT-PCR analysis of E-Cadherin (E-CAD), Vimentin (VIM), SNAIL and ZEB2 on OXDOs (orange bars) and MXDOs (blue bars). *P < 0.05, **P < 0.01 and ***P < 0.001 from two-tailed Student’s t test. (H) Invasion/migration assay performed with OXDOs and MXDOs. (I) Cell viability of OXDOs (orange bars) and MXDOs (blue bars) treated with 5 μM 5-Fluorouracil (5-FU) for 6 days. Values represent mean ± SD of three independent experiments. *P < 0.05 by unpaired Student’s t test.

Article Snippet: Vimentin (#5741), N-Cadherin (#13116), SLUG (#9585), ZEB1 (#3396), pSMAD 2/3 (#8828), TGFβ (#3711) were from Cell Signaling Technology, E-Cadherin (#610181) from Becton Dickinson, LTBP1 from Santa Cruz (#sc-271140).

Techniques: Western Blot, Control, Quantitative RT-PCR, Two Tailed Test, Staining, Activation Assay, Derivative Assay, Migration