Saline:Article Title: Polypharmacology‐Driven Discovery of ZAK‐I‐57: A Potent Multi‐Targeted Benzoxazinone Small Molecule for Hepatocellular Carcinoma Therapy
Article Snippet: .. Membranes were blocked with 5% non‐fat milk in Tris‐buffered saline Tween 20 (TBST) for 1 h. The blots were incubated with primary antibodies (EGFR [4267S, Cell Signaling Technology, MA, USA], c‐Myc [5605S, Cell Signaling Technology], Bax [2772S, Cell Signaling Technology], Bcl‐2 [sc‐7382, Santa Cruze, TX, USA], ERBB2 [2165S, Cell Signaling Technology], and α‐tubulin [62204, Invitrogen, MA, USA], 1:1000) at 4°C overnight. .. Subsequently, the membranes were washed thrice (15 min each) with TBST solution and incubated for 1 h with secondary horseradish peroxidase‐conjugated antibodies (NA934 and NA931, Cytiva, MA, USA, 1:5000).
Incubation:Article Title: Polypharmacology‐Driven Discovery of ZAK‐I‐57: A Potent Multi‐Targeted Benzoxazinone Small Molecule for Hepatocellular Carcinoma Therapy
Article Snippet: .. Membranes were blocked with 5% non‐fat milk in Tris‐buffered saline Tween 20 (TBST) for 1 h. The blots were incubated with primary antibodies (EGFR [4267S, Cell Signaling Technology, MA, USA], c‐Myc [5605S, Cell Signaling Technology], Bax [2772S, Cell Signaling Technology], Bcl‐2 [sc‐7382, Santa Cruze, TX, USA], ERBB2 [2165S, Cell Signaling Technology], and α‐tubulin [62204, Invitrogen, MA, USA], 1:1000) at 4°C overnight. .. Subsequently, the membranes were washed thrice (15 min each) with TBST solution and incubated for 1 h with secondary horseradish peroxidase‐conjugated antibodies (NA934 and NA931, Cytiva, MA, USA, 1:5000).
Western Blot:Article Title: N-phenylquinazolin-4-amine-based EGFR TKIs suppress pulmonary fibrosis by modulating the EGFR/ERBB3 axis in epithelial-macrophage interaction.
Article Snippet: For quantification, aminimumof five images were acquired per section, and positively stained areas were measured with ImageJ software (NIH, Bethesda, MD; http:// imagej.net/). .. Antibodies for immunoblotting, immunofluorescence, and immunohistochemistry staining Immunoblotting, IHC, and IF staining were performed using primary antibodies against p-EGFR (IHC 1:100; immunoblotting 1:1000; Cell Signalling; #2234S), EGFR(IHC 1:100; immunoblotting 1:1000; Santa Cruz Biotechnology; #sc102), p-ERBB2 (IHC 1:200; immunoblotting 1:1000; Novusbio; #NB100-89474), ERBB2 (IHC 1:100; immunoblotting 1:1000; Cell Signalling; #2165), p-ERBB3 (IHC 1:200; immunoblotting 1:2000; Abcam; #ab131444), ERBB3 (IHC 1:100; Invitrogen; #MA513008), p-ERBB4 (IHC 1:200; immunoblotting 1:1000; Abcam; #ab61059), ERBB4 (IHC 1:200; immunoblotting 1:1000; Abcam; #ab19391), pro-SPC (IF 1:200; Merck; #AB3786), T1a (IF 1:200; R&D System; AF3244), arginase-1 (IHC/IF 1:2000; Novusbio; #NBP1-32731), iNOS (IHC/IF 1:100; Santa Cruz Biotechnology; #sc7271), TNF-α (IHC/ IF 1:100; R&D System; #AF-410NA), IL-6 (IHC/IF 1:200; Abcam; #ab208113), IFN-γ (IHC/IF 1:200; Abcam; #ab216642), EGF (IHC/IF 1:100; Santa Cruz Biotechnology; #sc166779), EREG (IHC/IF 1:100; Santa Cruz Biotechnology; #sc376284), NRG1 (IHC/IF 1:200; Invitrogen; #MA5-12896), AREG (IHC/IF 1:200; R&D System; #AF989), FSP (IF 1:200; Novus; NBP2-54580AF488), Fibronectin (IHC1:100; Abcam; #ab2413), MMP9 (IHC; 1:100 santa Cruz Biothechnology #sc6841), tdTomato (IF1:10,000; LSbio; #LS-C340696), and β-actin (immunoblotting 1:1000; Abcam; #ab8226). .. Actin stressfibers in cells were stainedwith Alexa Fluor 546-conjugated phalloidin (1:200; Invitrogen; #A22283), which specifically binds F-actin.
Article Title: N-phenylquinazolin-4-amine-based EGFR TKIs suppress pulmonary fibrosis by modulating the EGFR/ERBB3 axis in epithelial–macrophage interaction
Article Snippet: .. Immunoblotting, IHC, and IF staining were performed using primary antibodies against p-EGFR (IHC 1:100; immunoblotting 1:1000; Cell Signalling; #2234S), EGFR(IHC 1:100; immunoblotting 1:1000; Santa Cruz Biotechnology; #sc102), p-ERBB2 (IHC 1:200; immunoblotting 1:1000; Novusbio; #NB100-89474), ERBB2 (IHC 1:100; immunoblotting 1:1000; Cell Signalling; #2165), p-ERBB3 (IHC 1:200; immunoblotting 1:2000; Abcam; #ab131444), ERBB3 (IHC 1:100; Invitrogen; #MA5-13008), p-ERBB4 (IHC 1:200; immunoblotting 1:1000; Abcam; #ab61059), ERBB4 (IHC 1:200; immunoblotting 1:1000; Abcam; #ab19391), pro-SPC (IF 1:200; Merck; #AB3786), T1a (IF 1:200; R&D System; AF3244), arginase-1 (IHC/IF 1:2000; Novusbio; #NBP1-32731), iNOS (IHC/IF 1:100; Santa Cruz Biotechnology; #sc7271), TNF-α (IHC/IF 1:100; R&D System; #AF-410NA), IL-6 (IHC/IF 1:200; Abcam; #ab208113), IFN-γ (IHC/IF 1:200; Abcam; #ab216642), EGF (IHC/IF 1:100; Santa Cruz Biotechnology; #sc166779), EREG (IHC/IF 1:100; Santa Cruz Biotechnology; #sc376284), NRG1 (IHC/IF 1:200; Invitrogen; #MA5-12896), AREG (IHC/IF 1:200; R&D System; #AF989), FSP (IF 1:200; Novus; NBP2-54580AF488), Fibronectin (IHC1:100; Abcam; #ab2413), MMP9 (IHC; 1:100 santa Cruz Biothechnology #sc6841), tdTomato (IF1:10,000; LSbio; #LS-C340696), and β-actin (immunoblotting 1:1000; Abcam; #ab8226). .. Actin stress fibers in cells were stained with Alexa Fluor 546-conjugated phalloidin (1:200; Invitrogen; #A22283), which specifically binds F-actin.
Immunofluorescence:Article Title: N-phenylquinazolin-4-amine-based EGFR TKIs suppress pulmonary fibrosis by modulating the EGFR/ERBB3 axis in epithelial-macrophage interaction.
Article Snippet: For quantification, aminimumof five images were acquired per section, and positively stained areas were measured with ImageJ software (NIH, Bethesda, MD; http:// imagej.net/). .. Antibodies for immunoblotting, immunofluorescence, and immunohistochemistry staining Immunoblotting, IHC, and IF staining were performed using primary antibodies against p-EGFR (IHC 1:100; immunoblotting 1:1000; Cell Signalling; #2234S), EGFR(IHC 1:100; immunoblotting 1:1000; Santa Cruz Biotechnology; #sc102), p-ERBB2 (IHC 1:200; immunoblotting 1:1000; Novusbio; #NB100-89474), ERBB2 (IHC 1:100; immunoblotting 1:1000; Cell Signalling; #2165), p-ERBB3 (IHC 1:200; immunoblotting 1:2000; Abcam; #ab131444), ERBB3 (IHC 1:100; Invitrogen; #MA513008), p-ERBB4 (IHC 1:200; immunoblotting 1:1000; Abcam; #ab61059), ERBB4 (IHC 1:200; immunoblotting 1:1000; Abcam; #ab19391), pro-SPC (IF 1:200; Merck; #AB3786), T1a (IF 1:200; R&D System; AF3244), arginase-1 (IHC/IF 1:2000; Novusbio; #NBP1-32731), iNOS (IHC/IF 1:100; Santa Cruz Biotechnology; #sc7271), TNF-α (IHC/ IF 1:100; R&D System; #AF-410NA), IL-6 (IHC/IF 1:200; Abcam; #ab208113), IFN-γ (IHC/IF 1:200; Abcam; #ab216642), EGF (IHC/IF 1:100; Santa Cruz Biotechnology; #sc166779), EREG (IHC/IF 1:100; Santa Cruz Biotechnology; #sc376284), NRG1 (IHC/IF 1:200; Invitrogen; #MA5-12896), AREG (IHC/IF 1:200; R&D System; #AF989), FSP (IF 1:200; Novus; NBP2-54580AF488), Fibronectin (IHC1:100; Abcam; #ab2413), MMP9 (IHC; 1:100 santa Cruz Biothechnology #sc6841), tdTomato (IF1:10,000; LSbio; #LS-C340696), and β-actin (immunoblotting 1:1000; Abcam; #ab8226). .. Actin stressfibers in cells were stainedwith Alexa Fluor 546-conjugated phalloidin (1:200; Invitrogen; #A22283), which specifically binds F-actin.
Immunohistochemistry:Article Title: N-phenylquinazolin-4-amine-based EGFR TKIs suppress pulmonary fibrosis by modulating the EGFR/ERBB3 axis in epithelial-macrophage interaction.
Article Snippet: For quantification, aminimumof five images were acquired per section, and positively stained areas were measured with ImageJ software (NIH, Bethesda, MD; http:// imagej.net/). .. Antibodies for immunoblotting, immunofluorescence, and immunohistochemistry staining Immunoblotting, IHC, and IF staining were performed using primary antibodies against p-EGFR (IHC 1:100; immunoblotting 1:1000; Cell Signalling; #2234S), EGFR(IHC 1:100; immunoblotting 1:1000; Santa Cruz Biotechnology; #sc102), p-ERBB2 (IHC 1:200; immunoblotting 1:1000; Novusbio; #NB100-89474), ERBB2 (IHC 1:100; immunoblotting 1:1000; Cell Signalling; #2165), p-ERBB3 (IHC 1:200; immunoblotting 1:2000; Abcam; #ab131444), ERBB3 (IHC 1:100; Invitrogen; #MA513008), p-ERBB4 (IHC 1:200; immunoblotting 1:1000; Abcam; #ab61059), ERBB4 (IHC 1:200; immunoblotting 1:1000; Abcam; #ab19391), pro-SPC (IF 1:200; Merck; #AB3786), T1a (IF 1:200; R&D System; AF3244), arginase-1 (IHC/IF 1:2000; Novusbio; #NBP1-32731), iNOS (IHC/IF 1:100; Santa Cruz Biotechnology; #sc7271), TNF-α (IHC/ IF 1:100; R&D System; #AF-410NA), IL-6 (IHC/IF 1:200; Abcam; #ab208113), IFN-γ (IHC/IF 1:200; Abcam; #ab216642), EGF (IHC/IF 1:100; Santa Cruz Biotechnology; #sc166779), EREG (IHC/IF 1:100; Santa Cruz Biotechnology; #sc376284), NRG1 (IHC/IF 1:200; Invitrogen; #MA5-12896), AREG (IHC/IF 1:200; R&D System; #AF989), FSP (IF 1:200; Novus; NBP2-54580AF488), Fibronectin (IHC1:100; Abcam; #ab2413), MMP9 (IHC; 1:100 santa Cruz Biothechnology #sc6841), tdTomato (IF1:10,000; LSbio; #LS-C340696), and β-actin (immunoblotting 1:1000; Abcam; #ab8226). .. Actin stressfibers in cells were stainedwith Alexa Fluor 546-conjugated phalloidin (1:200; Invitrogen; #A22283), which specifically binds F-actin.
Article Title: N-phenylquinazolin-4-amine-based EGFR TKIs suppress pulmonary fibrosis by modulating the EGFR/ERBB3 axis in epithelial–macrophage interaction
Article Snippet: .. Immunoblotting, IHC, and IF staining were performed using primary antibodies against p-EGFR (IHC 1:100; immunoblotting 1:1000; Cell Signalling; #2234S), EGFR(IHC 1:100; immunoblotting 1:1000; Santa Cruz Biotechnology; #sc102), p-ERBB2 (IHC 1:200; immunoblotting 1:1000; Novusbio; #NB100-89474), ERBB2 (IHC 1:100; immunoblotting 1:1000; Cell Signalling; #2165), p-ERBB3 (IHC 1:200; immunoblotting 1:2000; Abcam; #ab131444), ERBB3 (IHC 1:100; Invitrogen; #MA5-13008), p-ERBB4 (IHC 1:200; immunoblotting 1:1000; Abcam; #ab61059), ERBB4 (IHC 1:200; immunoblotting 1:1000; Abcam; #ab19391), pro-SPC (IF 1:200; Merck; #AB3786), T1a (IF 1:200; R&D System; AF3244), arginase-1 (IHC/IF 1:2000; Novusbio; #NBP1-32731), iNOS (IHC/IF 1:100; Santa Cruz Biotechnology; #sc7271), TNF-α (IHC/IF 1:100; R&D System; #AF-410NA), IL-6 (IHC/IF 1:200; Abcam; #ab208113), IFN-γ (IHC/IF 1:200; Abcam; #ab216642), EGF (IHC/IF 1:100; Santa Cruz Biotechnology; #sc166779), EREG (IHC/IF 1:100; Santa Cruz Biotechnology; #sc376284), NRG1 (IHC/IF 1:200; Invitrogen; #MA5-12896), AREG (IHC/IF 1:200; R&D System; #AF989), FSP (IF 1:200; Novus; NBP2-54580AF488), Fibronectin (IHC1:100; Abcam; #ab2413), MMP9 (IHC; 1:100 santa Cruz Biothechnology #sc6841), tdTomato (IF1:10,000; LSbio; #LS-C340696), and β-actin (immunoblotting 1:1000; Abcam; #ab8226). .. Actin stress fibers in cells were stained with Alexa Fluor 546-conjugated phalloidin (1:200; Invitrogen; #A22283), which specifically binds F-actin.
Staining:Article Title: N-phenylquinazolin-4-amine-based EGFR TKIs suppress pulmonary fibrosis by modulating the EGFR/ERBB3 axis in epithelial-macrophage interaction.
Article Snippet: For quantification, aminimumof five images were acquired per section, and positively stained areas were measured with ImageJ software (NIH, Bethesda, MD; http:// imagej.net/). .. Antibodies for immunoblotting, immunofluorescence, and immunohistochemistry staining Immunoblotting, IHC, and IF staining were performed using primary antibodies against p-EGFR (IHC 1:100; immunoblotting 1:1000; Cell Signalling; #2234S), EGFR(IHC 1:100; immunoblotting 1:1000; Santa Cruz Biotechnology; #sc102), p-ERBB2 (IHC 1:200; immunoblotting 1:1000; Novusbio; #NB100-89474), ERBB2 (IHC 1:100; immunoblotting 1:1000; Cell Signalling; #2165), p-ERBB3 (IHC 1:200; immunoblotting 1:2000; Abcam; #ab131444), ERBB3 (IHC 1:100; Invitrogen; #MA513008), p-ERBB4 (IHC 1:200; immunoblotting 1:1000; Abcam; #ab61059), ERBB4 (IHC 1:200; immunoblotting 1:1000; Abcam; #ab19391), pro-SPC (IF 1:200; Merck; #AB3786), T1a (IF 1:200; R&D System; AF3244), arginase-1 (IHC/IF 1:2000; Novusbio; #NBP1-32731), iNOS (IHC/IF 1:100; Santa Cruz Biotechnology; #sc7271), TNF-α (IHC/ IF 1:100; R&D System; #AF-410NA), IL-6 (IHC/IF 1:200; Abcam; #ab208113), IFN-γ (IHC/IF 1:200; Abcam; #ab216642), EGF (IHC/IF 1:100; Santa Cruz Biotechnology; #sc166779), EREG (IHC/IF 1:100; Santa Cruz Biotechnology; #sc376284), NRG1 (IHC/IF 1:200; Invitrogen; #MA5-12896), AREG (IHC/IF 1:200; R&D System; #AF989), FSP (IF 1:200; Novus; NBP2-54580AF488), Fibronectin (IHC1:100; Abcam; #ab2413), MMP9 (IHC; 1:100 santa Cruz Biothechnology #sc6841), tdTomato (IF1:10,000; LSbio; #LS-C340696), and β-actin (immunoblotting 1:1000; Abcam; #ab8226). .. Actin stressfibers in cells were stainedwith Alexa Fluor 546-conjugated phalloidin (1:200; Invitrogen; #A22283), which specifically binds F-actin.
Article Title: Recombinant Human Neuregulin1-β1 Significantly Reduces Schwannoma Growth in Mice.
Article Snippet: .. A total of 4μm thick cross sections were used for hematoxylin and eosin (H&E) staining and immunohistochemical labeling of S100beta (S100b) (1:8000; Dako; GA50461-2), myelin protein zero (MPZ) (1:300; Bioss Antibodies; bs-0337R), p75 (1:200; Millipore; AB1554), and ErbB2 (1:500; Cell Signaling; 2,165) in an automated Ventana stainer (Ventana Medical Systems, USA), using standard antigen retrieval protocols (CC1st, no pretreatment for S100b-protein). .. Immunoblotting was completed as described in Morrison et al,19 using the following primary antibodies: c-Jun (1:1,000; Cell Signaling; 8,752), anti-ErbB2 (1:500; Cell Signaling; 2,165), anti-GAPDH (1:1,000; Santa Cruz; 6C5), myelin basic protein (MBP) (1:500; Millipore; MAB384), and anti-merlin (1:500; Santa Cruz; A19).
Article Title: N-phenylquinazolin-4-amine-based EGFR TKIs suppress pulmonary fibrosis by modulating the EGFR/ERBB3 axis in epithelial–macrophage interaction
Article Snippet: .. Immunoblotting, IHC, and IF staining were performed using primary antibodies against p-EGFR (IHC 1:100; immunoblotting 1:1000; Cell Signalling; #2234S), EGFR(IHC 1:100; immunoblotting 1:1000; Santa Cruz Biotechnology; #sc102), p-ERBB2 (IHC 1:200; immunoblotting 1:1000; Novusbio; #NB100-89474), ERBB2 (IHC 1:100; immunoblotting 1:1000; Cell Signalling; #2165), p-ERBB3 (IHC 1:200; immunoblotting 1:2000; Abcam; #ab131444), ERBB3 (IHC 1:100; Invitrogen; #MA5-13008), p-ERBB4 (IHC 1:200; immunoblotting 1:1000; Abcam; #ab61059), ERBB4 (IHC 1:200; immunoblotting 1:1000; Abcam; #ab19391), pro-SPC (IF 1:200; Merck; #AB3786), T1a (IF 1:200; R&D System; AF3244), arginase-1 (IHC/IF 1:2000; Novusbio; #NBP1-32731), iNOS (IHC/IF 1:100; Santa Cruz Biotechnology; #sc7271), TNF-α (IHC/IF 1:100; R&D System; #AF-410NA), IL-6 (IHC/IF 1:200; Abcam; #ab208113), IFN-γ (IHC/IF 1:200; Abcam; #ab216642), EGF (IHC/IF 1:100; Santa Cruz Biotechnology; #sc166779), EREG (IHC/IF 1:100; Santa Cruz Biotechnology; #sc376284), NRG1 (IHC/IF 1:200; Invitrogen; #MA5-12896), AREG (IHC/IF 1:200; R&D System; #AF989), FSP (IF 1:200; Novus; NBP2-54580AF488), Fibronectin (IHC1:100; Abcam; #ab2413), MMP9 (IHC; 1:100 santa Cruz Biothechnology #sc6841), tdTomato (IF1:10,000; LSbio; #LS-C340696), and β-actin (immunoblotting 1:1000; Abcam; #ab8226). .. Actin stress fibers in cells were stained with Alexa Fluor 546-conjugated phalloidin (1:200; Invitrogen; #A22283), which specifically binds F-actin.
Immunohistochemical staining:Article Title: Recombinant Human Neuregulin1-β1 Significantly Reduces Schwannoma Growth in Mice.
Article Snippet: .. A total of 4μm thick cross sections were used for hematoxylin and eosin (H&E) staining and immunohistochemical labeling of S100beta (S100b) (1:8000; Dako; GA50461-2), myelin protein zero (MPZ) (1:300; Bioss Antibodies; bs-0337R), p75 (1:200; Millipore; AB1554), and ErbB2 (1:500; Cell Signaling; 2,165) in an automated Ventana stainer (Ventana Medical Systems, USA), using standard antigen retrieval protocols (CC1st, no pretreatment for S100b-protein). .. Immunoblotting was completed as described in Morrison et al,19 using the following primary antibodies: c-Jun (1:1,000; Cell Signaling; 8,752), anti-ErbB2 (1:500; Cell Signaling; 2,165), anti-GAPDH (1:1,000; Santa Cruz; 6C5), myelin basic protein (MBP) (1:500; Millipore; MAB384), and anti-merlin (1:500; Santa Cruz; A19).
Labeling:Article Title: Recombinant Human Neuregulin1-β1 Significantly Reduces Schwannoma Growth in Mice.
Article Snippet: .. A total of 4μm thick cross sections were used for hematoxylin and eosin (H&E) staining and immunohistochemical labeling of S100beta (S100b) (1:8000; Dako; GA50461-2), myelin protein zero (MPZ) (1:300; Bioss Antibodies; bs-0337R), p75 (1:200; Millipore; AB1554), and ErbB2 (1:500; Cell Signaling; 2,165) in an automated Ventana stainer (Ventana Medical Systems, USA), using standard antigen retrieval protocols (CC1st, no pretreatment for S100b-protein). .. Immunoblotting was completed as described in Morrison et al,19 using the following primary antibodies: c-Jun (1:1,000; Cell Signaling; 8,752), anti-ErbB2 (1:500; Cell Signaling; 2,165), anti-GAPDH (1:1,000; Santa Cruz; 6C5), myelin basic protein (MBP) (1:500; Millipore; MAB384), and anti-merlin (1:500; Santa Cruz; A19).
other:Article Title: Metformin enhances alpelisib sensitivity in HER2+ breast cancer by suppressing cancer stemness and oncogenic signaling
Article Snippet: Primary antibodies against phosphorylated ERK1/2 (Thr202/Tyr204) (p-ERK1/2, Cat: 9101), ERK1/2(Cat: 9102), phosphorylated Akt (Ser473) (p-Akt, Cat: 4060), Akt(Cat: 9272), phosphorylated Stat3 (Tyr705) (p-Stat3, Cat: 9145), Stat3 (Cat: 4904), Cyclin D1(Cat: 55506), c-Myc (Cat: 9402), phosphorylated ErbB2 (Tyr1221/1222) (p-ErbB2, Cat: 2243), ErbB2 (Cat: 2242), β-Catenin (Cat: 8480), Active β-Catenin (Cat: 19807), phosphorylated mTOR (Ser2448) (p-mTOR, Cat: 5536), mTOR (Cat: 2983), phosphorylated S6K (Thr389) (p-S6K, Cat: 9234), S6K (Cat: 9202), Nanog (Cat: 4903), KLF4 (Cat: 12173), Sox2 (Cat: 23064), AMPK (Cat: 2532), p-AMPK (Cat: 2535) along with Anti-mouse/rabbit HRP-linked secondary antibodies, were purchased from Cell Signaling Technology (Danvers, MA, USA).
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