rabbit antihuman fibronectin  (Bio-Rad)

Journal: International Journal of Molecular Sciences

Article Title: Alpha-Lipoic Acid Reduces Cell Growth, Inhibits Autophagy, and Counteracts Prostate Cancer Cell Migration and Invasion: Evidence from In Vitro Studies

doi: 10.3390/ijms242317111

Figure Lengend Snippet: ( a ) Western blotting analysis shows a downregulation of TGFβ-1 and Vimentin proteins expression in both LNCaP and DU-145 after ALA treatment for 48 h. The bars represent the mean ± SD of three experiments. Optical density was evaluated as the optical density and is represented as fold change for treated vs. untreated cells normalized for the loading control. * p < 0.05; ** p < 0.001; *** p < 0.0001 treated vs. untreated cells. GAPDH was used as loading control. ( b ) Immunoblotting showing pNF-kB protein expressions in LNCaP and DU-145 cells exposed for 48 h to ALA. GAPDH was used as a loading control. The bars represent the mean ± SD of three experiments in which band intensities were evaluated as the optical density and are represented as fold change for treated vs. untreated cells normalized for the loading control. * p < 0.05; ** p < 0.001 treated vs. untreated cells. ( c ) Immunofluorescence assay (IF) of fibronectin protein was assessed for both prostate cancer cell lines. Cell monolayers were seeded and then treated with ALA for 24 h. The image showed a reduction in the signals of the fibronectin after ALA treatment (third panels) in both cell lines compared to untreated cells. The bars represent the mean ± SD of three experiments. Mean fluorescence intensity is represented as fold change for treated vs. untreated cells. *** p < 0.0001. Scale bar: 12.5 μm.

Article Snippet: Prostate cancer cells were seeded into a six-well multiwell and synchronized in SFM for 12 h. Cells were exposed to ALA (271 μM for LNCaP; 278 μM for DU-145) for 24 h and then fixed with methanol followed by 5% bovine serum albumin blocking and incubated with a rabbit antihuman fibronectin antibody (sc-9068), Santa Cruz Biotechnology, Santa Cruz, CA, USA.

Techniques: Western Blot, Expressing, Control, Immunofluorescence, Fluorescence

Journal: Biomaterials advances

Article Title: The shape of our gut: Dissecting its impact on drug absorption in a 3D bioprinted intestinal model.

doi: 10.1016/j.bioadv.2023.213564

Figure Lengend Snippet: Fig. 3. Fibronectin (gray), vimentin (green), alpha-SMA (green) and collagen type IV (magenta) secretion by HIFs embedded in hydrogels (2.5 × 106 cells/mL) after 21 days in culture. Merged (left) and single channels (right). Fibroblasts presented an elongated shape and were able to produce and secrete ECM proteins into the hydrogel. Pictures depict a maximum intensity projection (z range 250–300 μm) (Scale bar = 50 μm).

Article Snippet: Mouse anti-human vimentin (1:50) (sc-6260, Santa Cruz Biotechnolgy) + rabbit antihuman fibronectin (1:100) (F3648, Sigma) or mouse anti-human alpha smooth muscle actin (alpha-SMA) (1:200) (ab7817, Abcam) + goat antihuman collagen type IV (1:250) (134001, Biorad) were diluted in the primary working buffer (PWB) containing 0.1 % BSA, 0.3 % donkey serum and 0.2 % Triton X-100 in PBS and incubated with the samples overnight (ON), at 4 ◦C in shaking conditions.

Techniques:

Journal: Biomaterials advances

Article Title: The shape of our gut: Dissecting its impact on drug absorption in a 3D bioprinted intestinal model.

doi: 10.1016/j.bioadv.2023.213564

Figure Lengend Snippet: Fig. 7. Protein expression of (a) β-catenin (red), (b) E-cadherin (gray) and fibronectin (yellow), (c) collagen type IV (magenta) and fibronectin (yellow) and (d) β-catenin (red) and ZO-1 (green) in the villi scaffolds after 21 days in culture. * denote the basolateral side of the epithelium (hydrogel). [(a) Scale bar = 100 μm; (b, c) Scale bar = 50 μm].

Article Snippet: Mouse anti-human vimentin (1:50) (sc-6260, Santa Cruz Biotechnolgy) + rabbit antihuman fibronectin (1:100) (F3648, Sigma) or mouse anti-human alpha smooth muscle actin (alpha-SMA) (1:200) (ab7817, Abcam) + goat antihuman collagen type IV (1:250) (134001, Biorad) were diluted in the primary working buffer (PWB) containing 0.1 % BSA, 0.3 % donkey serum and 0.2 % Triton X-100 in PBS and incubated with the samples overnight (ON), at 4 ◦C in shaking conditions.

Techniques: Expressing

Journal: Journal of Biomechanical Engineering

Article Title: The Effect of Cyclic Strain on Human Fibroblasts With Lamin A/C Mutations and Its Relation to Heart Disease

doi: 10.1115/1.4044091

Figure Lengend Snippet: Summary of sample sizes of statistically tested groups

Article Snippet: Once fixed, the cultures were stained for nuclei (4,6-Diamidino-2-Phenylindole Dihydrochloride, DAPI, ThermoFisher, Grand Island, NY), actin (Alexa Fluor 488 Phalloidin, ThermoFisher, Grand Island, NY), and fibronectin (polyclonal rabbit antihuman fibronectin, Sigma-Aldrich, Saint Louis, MO).

Techniques:

Journal: Journal of Biomechanical Engineering

Article Title: The Effect of Cyclic Strain on Human Fibroblasts With Lamin A/C Mutations and Its Relation to Heart Disease

doi: 10.1115/1.4044091

Figure Lengend Snippet: Summary of sample sizes of statistically tested groups for 72-h experiments

Article Snippet: Once fixed, the cultures were stained for nuclei (4,6-Diamidino-2-Phenylindole Dihydrochloride, DAPI, ThermoFisher, Grand Island, NY), actin (Alexa Fluor 488 Phalloidin, ThermoFisher, Grand Island, NY), and fibronectin (polyclonal rabbit antihuman fibronectin, Sigma-Aldrich, Saint Louis, MO).

Techniques:

Journal: Journal of Biomechanical Engineering

Article Title: The Effect of Cyclic Strain on Human Fibroblasts With Lamin A/C Mutations and Its Relation to Heart Disease

doi: 10.1115/1.4044091

Figure Lengend Snippet: Consequences of cyclic strain on cell viability/proliferation and organization. (a) (i–ii) Example images of nuclei stained with DAPI (blue (print: gray)); (iii) cell density estimated by nuclei count. (b) (i–ii) example images of actin stained with Phalloidin (green (print: gray)); (iii) quantification of actin organization. (c) (i–ii) Example images of samples stained for fibronectin (red (print: gray)); (iii) quantification of extracellular matrix organization. (a–c) Scale bar: 25 μm; (i) stains in the static condition; (ii) stains in the stretch condition with black arrows indicating the direction of stretch; (iii) connecting lines represent significances within conditions and between corresponding groups (p < 0.05); sample sizes listed within the bars. Summary of statistical analysis and sample sizes found in Table ​Table11.

Article Snippet: Once fixed, the cultures were stained for nuclei (4,6-Diamidino-2-Phenylindole Dihydrochloride, DAPI, ThermoFisher, Grand Island, NY), actin (Alexa Fluor 488 Phalloidin, ThermoFisher, Grand Island, NY), and fibronectin (polyclonal rabbit antihuman fibronectin, Sigma-Aldrich, Saint Louis, MO).

Techniques: Staining