Journal: Cancer Medicine

Article Title: LRP5 knockdown: effect on prostate cancer invasion growth and skeletal metastasis in vitro and in vivo

doi: 10.1002/cam4.111

Figure Lengend Snippet: Effect of DN-LRP5 on PC-3 cells invasion, colony formation, and migration in vitro . Control (PC-3-pcDNA3.1) and experimental (PC-3-DN-LRP5) cells invasive capacity was evaluated by using a Boyden chamber Matrigel invasion assay. After 18 h of incubation, the invaded cells were fixed, stained, and 10 random fields were counted. Number of cells invading is shown as bar diagram ± SEM (A) as described in “Material and Methods”. Using soft agar assay, the ability of control and experimental cells to form colonies was examined and the number of colonies formed in each group was counted (B) as described in “Material and Methods”. PC-3 cells migration was determined by wound healing assay by seeding PC-3-pcDNA3.1 and PC-3-DN-LRPP5 cells in six-well plates and allowing them to grow as a monolayer and making a wound as described in “Material and Methods”. Control and experimental migrating cells were photographed at different time points (C). Percent wound healing was recorded at different time points, and percentage of wound healing with respect to T 0 was calculated using the equation described in “Material and Methods”. Results are presented as the mean ± SEM of two different experiments in duplicate from control and experimental cells. Significant differences from the control is represented by an asterisk ( P < 0.05).

Article Snippet: The invasive capacity of 5 × 10 4 PC-3-pcDNA3.1 and PC-3-DN-LRP5 cells were examined using two-compartment Boyden chamber Matrigel invasion assay (Costar Transwell, Corning Corporation, Corning, NY) as described previously .

Techniques: Migration, In Vitro, Invasion Assay, Incubation, Staining, Soft Agar Assay, Wound Healing Assay

Journal:

Article Title: Dissection of Key Events in Tubular Epithelial to Myofibroblast Transition and Its Implications in Renal Interstitial Fibrosis

doi:

Figure Lengend Snippet: Functional disruption of the reconstituted TBM by the conditioned media from the transformed cells. Matrigels (15-μm depth) analogous to native TBM matrix were formed on the transwell filters and incubated with the conditioned media from either control or TGF-β1-treated HKC cells for 4 days. The integrity of Matrigels was assessed by bacterial translocation through the gels. A and B: Representative plates show the colonies formed from the bacteria passed through the Matrigels. C: Graphic presentation of the numbers of bacteria translocated via the gels. **, P < 0.01 versus control (n = 3).

Article Snippet: Bacterial translocation was performed for evaluating TBM integrity using a two-compartment Boyden chamber with transwell filters containing 3-μm diameter pores (Corning Co., Corning, NY).

Techniques: Functional Assay, Disruption, Transformation Assay, Incubation, Control, Translocation Assay, Bacteria

Journal:

Article Title: Dissection of Key Events in Tubular Epithelial to Myofibroblast Transition and Its Implications in Renal Interstitial Fibrosis

doi:

Figure Lengend Snippet: Increased invasive capacity of the transformed cells on Matrigel. HKC cells were seeded on the top of Matrigels in the transwell filters of the Boyden chamber and incubated with or without TGF-β1 for 2 and 5 days, respectively. The cell extensions that migrated across the Matrigels and passed through the pores of filters were counted after staining. A and B: Representative micrographs show the pores of the transwell filters filled with cell extensions after incubation without or with TGF-β1 for 5 days. C: Graphic presentation of the numbers of pores filled with cell extensions after incubation with or without TGF-β1 for 2 and 5 days, respectively. **, P < 0.01 versus control (n = 3).

Article Snippet: Bacterial translocation was performed for evaluating TBM integrity using a two-compartment Boyden chamber with transwell filters containing 3-μm diameter pores (Corning Co., Corning, NY).

Techniques: Transformation Assay, Incubation, Staining, Control