Journal: Stem Cell Research & Therapy

Article Title: Ang II-AT2R increases mesenchymal stem cell migration by signaling through the FAK and RhoA/Cdc42 pathways in vitro

doi: 10.1186/s13287-017-0617-z

Figure Lengend Snippet: Effect of different concentrations of Ang II on migration of human bone marrow MSCs. a Nondirectional migration ability of human bone marrow MSCs after stimulations with different concentrations of Ang II (10 –8 , 10 –7 , 10 –6 , 10 –5 , and 3 × 10 –5 M) examined using the scratch assay. Wound sites (areas cleared of cells in the center of the scratched area) were observed and photographed at 0 and 24 h (200×). b Quantitative results of wound healing. c Directional migration ability of human bone marrow MSCs after stimulations with the different concentrations of Ang II indicated examined using the Transwell migration assay. Migrated cells on the bottom surfaces of the Transwell inserts were stained with crystal violet and observed under a microscope (200×). d Quantitative results of cell migration. n = 3; * p < 0.05 vs normal. Ang II angiotensin II

Article Snippet: Modified Boyden chamber assays were conducted using Transwell polyester membrane filter inserts with 8-μm pores (Corning Inc., Corning, NY, USA) at a density of 500,000 cells/ml per Transwell (upper chamber) as described previously [ ].

Techniques: Migration, Wound Healing Assay, Transwell Migration Assay, Staining, Microscopy

Journal: Stem Cell Research & Therapy

Article Title: Ang II-AT2R increases mesenchymal stem cell migration by signaling through the FAK and RhoA/Cdc42 pathways in vitro

doi: 10.1186/s13287-017-0617-z

Figure Lengend Snippet: Effect of AT1R and AT2R antagonists on Ang II-mediated migration of MSCs. a Nondirectional migration ability of MSCs after stimulation with 100 nM Ang II following pretreatment with Losartan (5 μM) and/or PD-123319 (5 μM) examined with the scratch assay. Wound sites (areas cleared of cells in the center of the scratched area) were observed and photographed at 0 and 24 h (200×). b Quantitative results of wound healing. c Directional migration ability of MSCs after stimulation with 100 nM Ang II following pretreatment with Losartan (5 μM) and/or PD-123319 (5 μM) examined using the Transwell migration assay. Migrated cells on the bottom surfaces of the Transwell inserts were stained with crystal violet and observed under a microscope (200×). d Quantitative results of cell migration. Significant differences were found compared with the normal group ( n = 3; * p < 0.05) and compared with the control group ( n = 3; # p < 0.05). Ang II angiotensin II

Article Snippet: Modified Boyden chamber assays were conducted using Transwell polyester membrane filter inserts with 8-μm pores (Corning Inc., Corning, NY, USA) at a density of 500,000 cells/ml per Transwell (upper chamber) as described previously [ ].

Techniques: Migration, Wound Healing Assay, Transwell Migration Assay, Staining, Microscopy, Control

Journal: Stem Cell Research & Therapy

Article Title: Ang II-AT2R increases mesenchymal stem cell migration by signaling through the FAK and RhoA/Cdc42 pathways in vitro

doi: 10.1186/s13287-017-0617-z

Figure Lengend Snippet: Roles of FAK and Rho GTPases in the migration of MSCs. a Nondirectional migration ability of MSCs after stimulation with Ang II and/or inhibitors of FAK and the Rho GTPases examined using the scratch assay. Wound sites (areas cleared of cells in the center of the scratched area) were observed and photographed at 0 and 24 h (200×). b Quantitative results of wound healing. c Directional migration ability of MSCs after stimulation with Ang II and/or inhibitors of FAK and the Rho GTPases examined using the Transwell migration assay. Migrated cells on the bottom surfaces of the Transwell inserts were stained with crystal violet and observed under a microscope (200×). d Quantitative results of cell migration. n = 3; * p < 0.05 vs normal, # p < 0.05 vs the control group. Ang II angiotensin II

Article Snippet: Modified Boyden chamber assays were conducted using Transwell polyester membrane filter inserts with 8-μm pores (Corning Inc., Corning, NY, USA) at a density of 500,000 cells/ml per Transwell (upper chamber) as described previously [ ].

Techniques: Migration, Wound Healing Assay, Transwell Migration Assay, Staining, Microscopy, Control

Journal: Cancer Communications

Article Title: Crosstalk between IL‐15Rα + tumor‐associated macrophages and breast cancer cells reduces CD8 + T cell recruitment

doi: 10.1002/cac2.12311

Figure Lengend Snippet: Macrophages facilitate breast cancer resistance to IL‐15 therapy. (A) Mice were implanted with 4T1 breast tumor cells and treated with PBS or 3 μg IL‐15 (intravenously) on days 8 and 15. Tumor growth was analyzed by measuring volumes every 3 days. (B) Representative photographs of tumors resected 21 days after inoculation. (C) Tumor weight was measured. Each dot indicates one mouse ( n = 8). (D‐E) CFSE‐labeled CD8 + T cells sorted from tumor tissue were stimulated in vitro with 25 ng/mL IL‐15. (F) IFN‐γ levels of CD8 + T cell culture supernatant were measured by ELISA. (G) Killing assay of 4T1 tumor cells by CD8 + T cells in presence of IL‐15 after 48 h of coculture at different E:T ratios ( n = 6). (H) Raw264.7 macrophages coculture with 4T1 tumor cells and CD8 + T cells using transwell in the presence of IL‐15. Killing assay of CD8 + T cells against 4T1 tumor cells after 12 h of coculture at different E:T ratios ( n = 6). (I) Raw264.7 macrophages cocultured with tumor cells were sorted and then cocultured with CD8 + T cells in the presence of IL‐15. (J) TAMs sorted from tumor tissues were cocultured with CD8 + T cells in the presence of IL‐15. Proliferation of T cells was analyzed by flow cytometry. All values are expressed as the mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, and not significant (ns) by Student's t‐test. Abbreviations: IL‐15, Interleukin‐15; TAM, tumor‐associated macrophage; PBS, phosphate buffered saline; E:T, effector (CD8 + T cells):target (4T1 tumor cells); Co‐M, macrophage cocultured with tumor cells; CFSE, carboxyfluorescein diacetate succinimidyl ester; IFN‐γ, Interferon‐gamma; ELISA, enzyme‐linked immunosorbent assay; SEM, standard error of mean

Article Snippet: Raw264.7 cells or iBMDMs were co‐cultured with 4T1 tumor cells by using a Transwell culture system with 0.4 μm pore polyester membrane inserts (Corning HTS Transwell, Corning, NY, USA) for 3 days.

Techniques: Labeling, In Vitro, Cell Culture, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Saline

Journal: Microbiology Spectrum

Article Title: Prevalence and virulence potential of Aeromonas spp. isolated from human diarrheal samples in North East Italy

doi: 10.1128/spectrum.00807-23

Figure Lengend Snippet: Effect of Aeromonas strains isolated from patients with diarrhea on transepithelial electrical resistance (TEER) in Caco-2 cell monolayers. ( A and B ) Post-confluent Caco-2 monolayers grown on transwell inserts were exposed to Aeromonas strains, either Aer + or Aer − (MOI 1:20). After 2 h at 37°C culture medium was removed, the cells were washed and cultured in complete medium containing antibiotics. TEER values were measured after 5 h ( A ) and 24 h ( B ). TEER was expressed as a percentage of resistance normalized to untreated monolayers (100%). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001 versus untreated Caco-2. ( C ) Effect of the presence/absence of Aer gene for each testing Aeromonas isolates on TEER of Caco-2 monolayers. *** P < 0.001. ( D ) Immunofluorescence localization of tight junction proteins zonulin-1 (ZO-1) and occludin (OC). Caco-2 monolayers were treated as described above and stained with anti-ZO and anti-occludin 5 h later. Samples were visualized using an inverted confocal microscope.

Article Snippet: Caco-2 cells were seeded on Transwell polyester membrane cell culture inserts (transparent PET membrane: 1.0 cm 2 growth surface area, 0.4 μm pore size; BD Falcon) in 24-well plates and incubated with DMEM medium w/o antibiotics.

Techniques: Isolation, Cell Culture, Immunofluorescence, Staining, Microscopy