Journal: International journal of molecular sciences

Article Title: Preconditioned Chorionic Villus Mesenchymal Stem/Stromal Cells (CVMSCs) Minimize the Invasive Phenotypes of Breast Cancer Cell Line MDA231 In Vitro.

doi: 10.3390/ijms24119569

Figure Lengend Snippet: Figure 6. Modulation in expression of tumor suppressor proteins and oncogenes in MDA231 cells treated with preconditioned CVMSCs: flow cytometry analysis for the expression of tumor suppressor proteins modulated in MDA231 cells after treatment with preconditioned CVMSCs showed significant increase in the expression levels for CDH1 and IFN-gamma in both IC and CM settings as compared to untreated control (A(i)). Expression levels of oncogenes such as IDO, IL6, MMP7, and TGF-β1 reduced significantly in MDA231 cells in both IC and SF setting as compared to untreated control (B(i)). Data obtained by FACS analysis from three independent experiments were quantified and are shown in bar graphs as Mean Florescence Index (MFI) pertaining to CDH1 and IFN-gamma (A(ii)), and IDO, IL6, MMP7, and TGF-β1 (B(ii)), respectively. Bars represent standard errors. * p < 0.05.

Article Snippet: Fluorescent-labeled antibodies for flow cytometry experiments, including IFN-γ (Human IFN-gamma PE-conjugated Antibody) cat# IC285P; CDH1 (Human E-Cadherin PE-conjugated Antibody) cat# FAB18381P; IDO (Human Indoleamine 2,3-dioxygenase/IDO PE-conjugated Antibody) cat# IC6030P; IL6 (Human IL-6 PE-conjugated Antibody)cat# IC206P; MMP7 (Human MMP-7 PE-conjugated Antibody) cat# IC9071P; and TGF-β1 (Human TGF-beta 1 Alexa Fluor® 488-conjugated Antibody) cat# IC10502G were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing, Cytometry, Control

Journal: Cancer Medicine

Article Title: Role of Ca 2+ ‐Dependent Epithelial‐Mesenchymal Transition in Malignant Progression of Colorectal Cancer: Special Focus on REG Iα/ EDNRB

doi: 10.1002/cam4.71754

Figure Lengend Snippet: REG Iα promotes cell migration, invasion, and EMT via EDNRB. (A) Cell migration was assessed by Transwell assay. (B) Matrigel‐coated Transwell assay assessed cell invasion. (C) Western blot analysis of EMT‐related markers, including the epithelial marker E‐Cadherin and mesenchymal markers N‐Cadherin and Vimentin. β‐actin was used as the loading control. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: CHOP (Abcam, ab317378), Cleaved‐Caspase 3 (Abcam, ab32042), Cleaved‐PARP (Cell Signaling Technology, 9541), E‐Cadherin (BOSTER, PB9561), N‐Cadherin (BOSTER, A01577‐3), and Vimentin (BOSTER, BM4029).

Techniques: Migration, Transwell Assay, Western Blot, Marker, Control

Journal: Cancer Medicine

Article Title: Role of Ca 2+ ‐Dependent Epithelial‐Mesenchymal Transition in Malignant Progression of Colorectal Cancer: Special Focus on REG Iα/ EDNRB

doi: 10.1002/cam4.71754

Figure Lengend Snippet: The REG Iα‐EDNRB axis promotes cell migration, invasion, and EMT via the Ca 2+ signaling pathway. (A) Transwell assay assessed cell migration. (B) Cell invasion was assessed by Matrigel‐coated Transwell assay. (C) EMT‐related protein levels (E‐Cadherin, N‐Cadherin, Vimentin) were detected by Western blot. β‐actin served as the loading control. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: CHOP (Abcam, ab317378), Cleaved‐Caspase 3 (Abcam, ab32042), Cleaved‐PARP (Cell Signaling Technology, 9541), E‐Cadherin (BOSTER, PB9561), N‐Cadherin (BOSTER, A01577‐3), and Vimentin (BOSTER, BM4029).

Techniques: Migration, Transwell Assay, Western Blot, Control

Journal: Cancer Medicine

Article Title: Role of Ca 2+ ‐Dependent Epithelial‐Mesenchymal Transition in Malignant Progression of Colorectal Cancer: Special Focus on REG Iα/ EDNRB

doi: 10.1002/cam4.71754

Figure Lengend Snippet: The REG Iα‐EDNRB‐Ca 2+ axis promotes tumor growth and EMT in vivo. (A) Representative images of the xenograft tumors and excised tumor tissues from the indicated groups. (B) Statistical analysis of the final tumor weights. (C) Tumor growth curves measuring tumor volume over time. (D) H&E staining showed pathological changes in tumor tissues. (E) TUNEL assay detected cell apoptosis in tumor tissues. (F) Cell proliferation in tumor tissues was shown by Ki67 IHC staining. (G) Expression of REG Iα (Immunofluorescence, upper row) and EDNRB (IHC, lower row) in tumor tissues. (H) Western blot analysis of REG Iα, EDNRB, and p‐CaMKII protein levels in tumor tissues. β‐actin was used as the loading control. (I) Western blot analysis of EMT‐related proteins (E‐Cadherin, N‐Cadherin, Vimentin) in tumor tissues. β‐actin was used as the loading control. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: CHOP (Abcam, ab317378), Cleaved‐Caspase 3 (Abcam, ab32042), Cleaved‐PARP (Cell Signaling Technology, 9541), E‐Cadherin (BOSTER, PB9561), N‐Cadherin (BOSTER, A01577‐3), and Vimentin (BOSTER, BM4029).

Techniques: In Vivo, Staining, TUNEL Assay, Immunohistochemistry, Expressing, Immunofluorescence, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: Depletion of Lipocalin 2 (LCN2) in Mice Leads to Dysbiosis and Persistent Colonization with Segmented Filamentous Bacteria

doi: 10.3390/ijms222313156

Figure Lengend Snippet: Comparative analysis of the intestine macroscopical parameters and expression of Lcn2 in wild type (n = 4) and Lcn2 -/- mice (n = 4 or 5). ( A ) Evaluation of the body weight of male animals at the same age (left). Corresponding ceca were prepared and weighed (middle). Ratio of body to cecal weight (right). ( B ) Gross macroscopical and graphical comparison of the total small and large intestinal lengths measured (n = 4 or 5). ( C ) Quantitative analysis of Lcn2 mRNA (upper) and LCN2 protein expression (lower) in the distal small intestine (left) cecum (middle), and distal colon (right) of wild type ( Lcn2 +/+ ) and Lcn2 null mice (each n = 4 or 5). Relative mRNA expression of respective genes was done by RT-qPCR and normalized to Gapdh or β-actin . Equal protein loading was demonstrated by re-probing the membranes with a specific antibody directed against GAPDH or β-actin. E-cadherin was probed as a marker for epithelial-to-mesenchymal transition (EMT). Primers used are given in . Statistical evaluation in ( A – C ) was performed by an unpaired t -test, whereby the difference of two samples from a level of p < 0.05 is considered statistically significant (*: p < 0.05, **: p < 0.01, ***: p < 0.001).

Article Snippet: For detection of individual proteins, the primary antibodies for LCN2/NGAL (AF3508, R & D Systems, Abingdon, UK), GAPDH (6C5) (sc-32233, Santa Cruz, Biotech, Santa Cruz, CA, USA), β-actin (A5441, Sigma–Aldrich, MI, USA), and E-cadherin (AP20195PU-N, OriGene, Rockville, MD, USA) were diluted in 2.5% ( w / v ) non-fat milk powder in TBST.

Techniques: Expressing, Comparison, Quantitative RT-PCR, Marker

Journal: Scientific Reports

Article Title: Evaluating the origin and virulence of a Helicobacter pylori cagA -positive strain isolated from a non-human primate

doi: 10.1038/s41598-018-34425-4

Figure Lengend Snippet: Impact of H . pylori infection on ES cell-derived gastric organoids. ( a ) HE staining of ES-cell derived gastric organoids infected with H . pylori for 12 hours. Scale bar: 200 µm. ( b ) Immunostaining of gastric organoids infected with H . pylori for 12 hours. “No infection” indicates that the organoid was injected with bacteria-free Brucella Broth. HP , H . pylori ; E-cad, E-cadherin. Scale bar: 10 µm.

Article Snippet: The primary antibodies used for immunostaining are: α-SMA, mouse, Sigma, A5228, 1:100; β-Catenin, rabbit, SantaCruz, sc7199, 1:100; CagA, rabbit, AUSTRAL Biologicals, HPP5003-9, 1:100; E-cadherin, mouse, BD Biosciences, 610182, 1:100; E-cadherin, goat, R&D Systems, AF648, 1:100; Gastrin, mouse, Santa Cruz, sc28302, 1:100; H . pylori , rabbit, Abcam, ab80519, 1:100; Muc5AC, mouse, Abcam, ab3649, 1:100; Muc6, mouse, Abcam, Ab49462, 1:100; Nanog, rabbit, Abcam, ab21624, 1:500; Oct3/4, mouse, Santa Cruz, sc5279, 1:500; Vimentin, goat, Santa Cruz, sc7557, 1:200; ZO-1, rabbit, Invitrogen, 617300, 1:100.

Techniques: Infection, Derivative Assay, Staining, Immunostaining, Injection, Bacteria