Journal: Biochemistry and Biophysics Reports

Article Title: Integrin β1 subunit regulates cellular and secreted MUC5AC and MUC5B production in NCI–H292 human lung epithelial cells

doi: 10.1016/j.bbrep.2021.101124

Figure Lengend Snippet: Effects of integrin β1 overexpression on cellular and secreted mucin 5AC (MUC5AC) and mucin 5B (MUC5B) levels in NCI–H292 cells. A, NCI–H292 cells were transfected with integrin β1 (β1 cDNA) or control vectors (CNTL) and cultured on a 96-well plate. Integrin β1 and β-actin protein levels were detected in cells via immunoblotting with specific antibodies, and their levels were measured using ChemiDoc. B, MUC5AC levels in transfected cells were detected using the dot blot method and measured (cellular). C, MUC5AC levels were detected in the culture medium of transfected cells via the dot blot method and measured (secreted). Fold changes were based on control cells (mean ± standard deviation [SD], n = 5). D, MUC5B levels in the transfected cells were detected using the dot blot method and measured (cellular). E, MUC5B levels in the culture medium of the transfected cells were detected using the dot blot method and measured (secreted). Fold changes were based on control cells (mean ± SD, n = 5). Fold changes were normalized for cell numbers. Student's t-test was used to obtain p -values. Asterisks indicate statistical significance, * p < 0.05. Representative results for three independent experiments are presented.

Article Snippet: A, NCI–H292 cells were transfected with integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), and cultured on a 96-well plate.

Techniques: Over Expression, Transfection, Control, Cell Culture, Western Blot, Dot Blot, Standard Deviation

Journal: Biochemistry and Biophysics Reports

Article Title: Integrin β1 subunit regulates cellular and secreted MUC5AC and MUC5B production in NCI–H292 human lung epithelial cells

doi: 10.1016/j.bbrep.2021.101124

Figure Lengend Snippet: Effects of integrin α2 overexpression on cellular and secreted mucin 5AC (MUC5AC) levels in NCI–H292 cells. A, NCI–H292 cells were transfected with integrin α2 (α2 cDNA) or control vectors (CNTL) and cultured on a 96-well plate. A, MUC5AC levels in transfected cells were detected using the dot blot method and measured (cellular). B, MUC5AC levels were detected in the culture medium of transfected cells via the dot blot method and measured (secreted). Fold changes were based on control cells (mean ± standard deviation [SD], n = 5). Fold changes were based on control cells (mean ± SD, n = 5). Fold changes were normalized for cell numbers. Student's t-test was used to obtain p -values. Asterisks indicate statistical significance, * p < 0.05. Representative results for three independent experiments are presented.

Article Snippet: A, NCI–H292 cells were transfected with integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), and cultured on a 96-well plate.

Techniques: Over Expression, Transfection, Control, Cell Culture, Dot Blot, Standard Deviation

Journal: Biochemistry and Biophysics Reports

Article Title: Integrin β1 subunit regulates cellular and secreted MUC5AC and MUC5B production in NCI–H292 human lung epithelial cells

doi: 10.1016/j.bbrep.2021.101124

Figure Lengend Snippet: Effects of integrin β1 depletion on cellular and secreted mucin 5AC (MUC5AC) and ROS levels in NCI–H292 cells. A, NCI–H292 cells were transfected with integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), and cultured on a 96-well plate. Integrin β1 and β-actin protein levels were detected via immunoblotting and measured using ChemiDoc. B, MUC5AC levels in the integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), transfected cells were detected using the dot blot method and measured (cellular). C, MUC5AC levels in the culture medium of the in the integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), transfected cells were detected via the dot blot method and measured (secreted). D, MUC5AC levels in the integrin β1 siRNA (β1 siRNA, Thermo Fisher) or control siRNA (CNTL), transfected cells were detected using the dot blot method and measured (cellular). E, MUC5AC levels in the culture medium of the integrin β1 siRNA (β1 siRNA, Thermo Fisher) or control siRNA (CNTL), transfected cells were detected via the dot blot method and measured (secreted). F, ROS levels in the integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), transfected cells were detected by hydrogen peroxide/peroxidase assay (cellular). Fold changes were based on control cells (mean ± standard deviation, n = 5). Fold changes were normalized for cell numbers. Student's t-test was used to obtain p -values. Asterisks indicate statistical significance, * p < 0.05. Representative results for three independent experiments are presented.

Article Snippet: A, NCI–H292 cells were transfected with integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), and cultured on a 96-well plate.

Techniques: Transfection, Control, Cell Culture, Western Blot, Dot Blot, Standard Deviation

Journal: Biochemistry and Biophysics Reports

Article Title: Integrin β1 subunit regulates cellular and secreted MUC5AC and MUC5B production in NCI–H292 human lung epithelial cells

doi: 10.1016/j.bbrep.2021.101124

Figure Lengend Snippet: Effects of integrin β1 depletion on cellular and secreted mucin 5B (MUC5B) levels in NCI–H292 cells. A, MUC5B levels in the integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL) transfected cells were detected via the dot blot method and measured (cellular). B, MUC5B levels in the culture medium of the integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), transfected cells were detected via the dot blot method and measured (secreted). C, MUC5B levels in the integrin β1 siRNA (β1 siRNA, Thermo Fisher) or control siRNA (CNTL), transfected cells were detected via the dot blot method and measured (cellular). D, MUC5B levels in the culture medium of the integrin β1 siRNA (β1 siRNA, Thermo Fisher) or control siRNA (CNTL), transfected cells were detected via the dot blot method and measured (secreted). Fold changes were based on control cells (mean ± standard deviation, n = 5). Fold changes were normalized for cell numbers. Student's t-test was used to obtain p -values. Asterisks indicate statistical significance, * p < 0.05. Representative results for three independent experiments are presented.

Article Snippet: A, NCI–H292 cells were transfected with integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), and cultured on a 96-well plate.

Techniques: Control, Transfection, Dot Blot, Standard Deviation

Journal: Biochemistry and Biophysics Reports

Article Title: Integrin β1 subunit regulates cellular and secreted MUC5AC and MUC5B production in NCI–H292 human lung epithelial cells

doi: 10.1016/j.bbrep.2021.101124

Figure Lengend Snippet: Effects of integrin β1 overexpression or depletion on cellular mucin 5AC (MUC5AC) and mucin 5B (MUC5B) levels in NCI–H292 cells. NCI–H292 cells (3.1 × 10 4 cells/cm 2 ) were transfected with integrin β1 vector (β1 cDNA), control vector (CNTL), integrin β1 siRNA (β1 siRNA), or control siRNA (CNTL), and cultured on Lab-Tek Chamber Slides for 48 h MUC5AC and MUC5B protein expression was detected via immunohistochemical analysis (green). Cell nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI, blue). Yellow bars indicate 100 μm. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: A, NCI–H292 cells were transfected with integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), and cultured on a 96-well plate.

Techniques: Over Expression, Transfection, Plasmid Preparation, Control, Cell Culture, Expressing, Immunohistochemical staining, Staining

Journal: Biochemistry and Biophysics Reports

Article Title: Integrin β1 subunit regulates cellular and secreted MUC5AC and MUC5B production in NCI–H292 human lung epithelial cells

doi: 10.1016/j.bbrep.2021.101124

Figure Lengend Snippet: Activities of extracellular signal-regulated kinase (ERK) in NCI–H292 cells transfected with integrin β1 cDNA (β1 cDNA) or siRNA (β1 siRNA). A, NCI–H292 cells (1 × 10 4 cells/well) were transfected with integrin β1 cDNA or siRNA, cultured, and sampled in a 96-well plate. Phosphorylated ERK and total ERK protein levels were detected. Representative results for three independent experiments are presented. B, A densitometry analysis of phosphorylated ERK/total ERK in NCI–H292 cells which were transfected with integrin β1 cDNA or siRNA. C, D, NCI–H292 cells were transfected with integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology), control siRNA (CNTL) and cultured with (+) or without (−) 10 μM of U0126 on a 96-well plate. The cellular (cellular, C) and secreted (secreted, D) MUC5AC levels were detected via the dot blot method and measured.

Article Snippet: A, NCI–H292 cells were transfected with integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), and cultured on a 96-well plate.

Techniques: Transfection, Cell Culture, Control, Dot Blot