In Vitro:Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway
Article Snippet: The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.. The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.. The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain (1:2000; Molecular Probes, Eugene, OR, USA).The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain..
Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway.
Article Snippet: The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.. The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.. The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain (1:2000; Molecular Probes, Eugene, OR, USA).The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain..
Immunofluorescence:Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway
Article Snippet: The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.. The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.. The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain (1:2000; Molecular Probes, Eugene, OR, USA).The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain..
Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway.
Article Snippet: The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.. The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.. The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain (1:2000; Molecular Probes, Eugene, OR, USA).The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain..
Staining:Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway
Article Snippet: The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.. The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.. The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain (1:2000; Molecular Probes, Eugene, OR, USA).The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain..
Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway.
Article Snippet: The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.. The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.. The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain (1:2000; Molecular Probes, Eugene, OR, USA).The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain..
CCK-8 Assay:Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway
Article Snippet: The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.. The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.. The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain (1:2000; Molecular Probes, Eugene, OR, USA).The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain..
Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway.
Article Snippet: The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.. The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.. The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain (1:2000; Molecular Probes, Eugene, OR, USA).The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain..
Viability Assay:Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway
Article Snippet: The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.. The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.. The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain (1:2000; Molecular Probes, Eugene, OR, USA).The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain..
Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway.
Article Snippet: The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.. The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.. The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain (1:2000; Molecular Probes, Eugene, OR, USA).The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain..
BrdU Incorporation Assay:Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway
Article Snippet: The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.. The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.. The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain (1:2000; Molecular Probes, Eugene, OR, USA).The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain..
Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway.
Article Snippet: The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.. The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.. The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain (1:2000; Molecular Probes, Eugene, OR, USA).The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain..
LDH Cytotoxicity Assay:Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway
Article Snippet: The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.. The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.. The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain (1:2000; Molecular Probes, Eugene, OR, USA).The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain..
Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway.
Article Snippet: The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.. The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.. The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain (1:2000; Molecular Probes, Eugene, OR, USA).The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain..
Cell Cycle Assay:Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway
Article Snippet: The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.. The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.. The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain (1:2000; Molecular Probes, Eugene, OR, USA).The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain..
Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway.
Article Snippet: The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.. The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.. The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain (1:2000; Molecular Probes, Eugene, OR, USA).The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain..
Wound Healing Assay:Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway
Article Snippet: The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.. The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.. The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain (1:2000; Molecular Probes, Eugene, OR, USA).The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain..
Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway.
Article Snippet: The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.. The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.. The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain (1:2000; Molecular Probes, Eugene, OR, USA).The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain..
Western Blot:Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway
Article Snippet: The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.. The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.. The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain (1:2000; Molecular Probes, Eugene, OR, USA).The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain..
Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway.
Article Snippet: The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.The cells were permeabilized for 10 min with 0.5% Triton X-100 in PBS and blocked for 1 h with 5% skim milk in PBS at 25 °C.. The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.. The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain (1:2000; Molecular Probes, Eugene, OR, USA).The cells were treated with either fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG antibody or rabbit anti-goat antibody (1:100) for 2 h at 37 °C in the dark, and followed by counterstain with Hoechst 33,342 nuclear stain..
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