Journal: Journal of Carcinogenesis

Article Title: Clinicopathological significance of immunohistochemical expression of Filamin A in breast cancer

doi: 10.4103/jcar.JCar_9_20

Figure Lengend Snippet: Association of Filamin A expression with clinicopathological parameters

Article Snippet: Four-micron thick sections are cut from paraffin-embedded tissue blocks and were subjected to immunohistochemistry with anti-Filamin A/FLNa Picoband rabbit IgG polyclonal antibody (Boster Biological Technology, Pleasanton, CA, USA) in lyophilized form at dilution of 0.5 μg/ml with antibody diluent.

Techniques: Expressing

Journal: Journal of Carcinogenesis

Article Title: Clinicopathological significance of immunohistochemical expression of Filamin A in breast cancer

doi: 10.4103/jcar.JCar_9_20

Figure Lengend Snippet: Left: Photomicrograph showing Filamin A expressivity in nonneoplastic breast tissue showing membranous and cytoplasmic positivity (×10). Right: Photomicrograph showing a membranous pattern in myoepithelial cells in fibroadenoma (×10)

Article Snippet: Four-micron thick sections are cut from paraffin-embedded tissue blocks and were subjected to immunohistochemistry with anti-Filamin A/FLNa Picoband rabbit IgG polyclonal antibody (Boster Biological Technology, Pleasanton, CA, USA) in lyophilized form at dilution of 0.5 μg/ml with antibody diluent.

Techniques:

Journal: Journal of Carcinogenesis

Article Title: Clinicopathological significance of immunohistochemical expression of Filamin A in breast cancer

doi: 10.4103/jcar.JCar_9_20

Figure Lengend Snippet: Left: Photomicrograph showing invasive carcinoma of the breast (H and E, ×40). Right: Immunohistochemistry: Score 0 (absent) (×40) photomicrograph showing the absence of Filamin A expression in malignant cells

Article Snippet: Four-micron thick sections are cut from paraffin-embedded tissue blocks and were subjected to immunohistochemistry with anti-Filamin A/FLNa Picoband rabbit IgG polyclonal antibody (Boster Biological Technology, Pleasanton, CA, USA) in lyophilized form at dilution of 0.5 μg/ml with antibody diluent.

Techniques: Immunohistochemistry, Expressing

Journal: Journal of Carcinogenesis

Article Title: Clinicopathological significance of immunohistochemical expression of Filamin A in breast cancer

doi: 10.4103/jcar.JCar_9_20

Figure Lengend Snippet: Left: Photomicrograph showing invasive carcinoma of the breast (H and E, ×40). Right: Immunohistochemistry: Score 3 (strong positivity) (×40); photomicrograph showing strong cytoplasmic positivity of Filamin A expression in malignant cells

Article Snippet: Four-micron thick sections are cut from paraffin-embedded tissue blocks and were subjected to immunohistochemistry with anti-Filamin A/FLNa Picoband rabbit IgG polyclonal antibody (Boster Biological Technology, Pleasanton, CA, USA) in lyophilized form at dilution of 0.5 μg/ml with antibody diluent.

Techniques: Immunohistochemistry, Expressing

Journal: Advanced Science

Article Title: Nuclear Actin Polymerization Regulates Cell Epithelial‐Mesenchymal Transition

doi: 10.1002/advs.202300425

Figure Lengend Snippet: F/G‐actin binds nuclear proteins. A) Upper, HEK 293T cells were cultured in 0.5 µ m Jasp, 1 µ m LatB, or 10 µ m CytD for 1 h, lysed with lysis and F‐actin stabilization buffer 2 (LAS2), and subjected to F/G‐actin fractionation. Western blot showed that Jasp induced actin polymerization, while LatB and CytD induced actin depolymerization. Middle, The above cells were also resuspended in the fractionation buffer, and subjected to nuclear fractionation and F/G‐actin fractionation. Western blot showed that Jasp induced nuclear actin polymerization, while LatB and CytD treatment induced nuclear actin depolymerization. Lower, After cultured in 0.25 µ m Jasp, 0.5 µ m LatB, or 5 µ m CytD for 4 h, HEK 293T cells were subjected to nuclear fractionation. Nuclear extracts were lysed with LAS2, and subjected to immunoprecipitation with antibody against actin. The pulldown products were subjected to mass spectrometry analysis, showing that precipitation of nuclear actin pulled down α‐catenin, β‐catenin, and filamin A (FLNA) in Jasp treated cells, and MYBBP1A, NKRF, and profilin 1 (PFN1) in LatB and CytD treated cells. B) The cells were transfected with α‐catenin siRNAs. Western blot showed that silencing α‐catenin decreased α‐catenin expression, but did not affect β‐catenin expression. Silencing α‐catenin decreased both α‐catenin and β‐catenin in the nuclei. The nuclear extracts were incubated with 25 µL biotin‐X Phalloidin, and subjected to biotin‐labeled Phalloidin pulldown assay. Precipitation of F‐actin pulled down α‐catenin and β‐catenin, and pulled down less α‐catenin (23.4%) and β‐catenin (39.8%) by silencing α‐catenin. C) Western blot showed that silencing FLNA decreased its expression, but did not change SMAD2/SMAD3 levels. Silencing FLNA decreased FLNA, SMAD2, and SMAD3 in the nuclei. Precipitation of nuclear F‐actin pulled down FLNA, SMAD2, and SMAD3, but less FLNA (25.7%), SMAD2 (27.8%) and SMAD3 (10.5%) by silencing FLNA. D) Silencing PFN1 decreased PFN1 and MYPOP levels in cell lysate and nuclei. Precipitation of G‐actin pulled down PFN1 and MYPOP, but less of them by silencing PFN1. E) The nuclear fractions of the above cells were lysed with LAS2 and subjected to F/G‐actin fractionation and immunoprecipitation with antibody against actin. Precipitation of nuclear F‐actin pulled down β‐catenin, SMAD2, SMAD3, and precipitation of nuclear G‐actin pulled down MYBBP1A, NKRF, and MYPOP. F, HEK 293T cells were cultured in 0.1 µ m Jasp, 0.2 µ m LatB, and 1 µ m CytD for 16 h, lysed with lysis buffer, and subjected to Western blot. Jasp treatment repressed E‐cadherin, enhanced N‐cadherin and vimentin proteins. LatB and CytD treatment increased E‐cadherin, decreased N‐cadherin and vimentin levels.

Article Snippet: Monoclonal antibodies against FLNA (A3738) and profilin‐1 (A1164), and polyclonal antibodies against Myb‐binding protein 1A (MYBBP1A, A4429), NF‐kappa‐B‐repressing factor (NKRF, A4853), E‐cadherin (A3044), N‐cadherin (A0432), and vimentin (A11952) were purchased from ABclonal (Woburn, MA, USA).

Techniques: Cell Culture, Lysis, Fractionation, Western Blot, Immunoprecipitation, Mass Spectrometry, Transfection, Expressing, Incubation, Labeling