Journal: bioRxiv

Article Title: L-Arginine supplementation modulates L-Arg/NO metabolic processes and AMPK/ACC-1 signalling in BNL CL2 hepatocytes

doi: 10.64898/2026.02.03.703662

Figure Lengend Snippet: Cell growth/viable cell number ( a ) and culture viability ( b ) profiles of BNL CL2 cells with different concentrations of L-Arg (0, 400 and 800 µM) and the control complete DMEM media at 24, 48, 72 and 120 h. Relative mRNA expression of AMPK ( c ) and ACC-1 ( d ) cultured in different concentrations of L-Arg in BNL CL2 cells at 24 and 72 h. Data points represent the mean ± SD of each culture sample. Error bars represent the standard deviation from the mean (n = 3).

Article Snippet: The BNL CL2 cell line was used as a model system, which was a generous gift from Associate Prof of Medicine Dimiter Avtanski, Zucker School of Medicine at Hofstra/Northwell, Hempstead, New York.

Techniques: Control, Expressing, Cell Culture, Standard Deviation

Journal: bioRxiv

Article Title: L-Arginine supplementation modulates L-Arg/NO metabolic processes and AMPK/ACC-1 signalling in BNL CL2 hepatocytes

doi: 10.64898/2026.02.03.703662

Figure Lengend Snippet: Relative mRNA transcript expression (ΔΔCt) of AMPK (a) and ACC-1 (b) in the BNL CL2 cells with nitric oxide synthase inhibitor; L-NAME (4 mM) 24 and 72 h after addition and relative mRNA transcript expression of AMPK (c) and ACC-1 (d) in the BNL CL2 cells with nitric oxide donor; SNAP (100 µM) 6 and 24 h after addition. All these analyses were undertaken in BNL CL2 cells cultured in medium of different L-Arg concentrations (0, 400 and 800 µM) and control complete DMEM media. Data points represent the mean ± SD of each sample. Error bars represent the standard deviation from the mean (n =3).

Article Snippet: The BNL CL2 cell line was used as a model system, which was a generous gift from Associate Prof of Medicine Dimiter Avtanski, Zucker School of Medicine at Hofstra/Northwell, Hempstead, New York.

Techniques: Expressing, Cell Culture, Control, Standard Deviation

Journal: bioRxiv

Article Title: L-Arginine supplementation modulates L-Arg/NO metabolic processes and AMPK/ACC-1 signalling in BNL CL2 hepatocytes

doi: 10.64898/2026.02.03.703662

Figure Lengend Snippet: Western blot comparison of the expression of AMPK and ACC-1 proteins (a, c, e and g) involved in L-Arg/NO metabolic pathway signalling, and the amount of phospho-protein (b, d, f and h) of these targets in BNL CL2 cells cultured in no L-Arg SILAC DMEM media, control complete DMEM media and 400 and 800 µM L-Arg in L-Arg free SILAC DMEM media for 24 and 72 h and in one of the control, untreated culture samples at T=0. β-actin is used as a loading control. The same amount of protein (10 µg) from different treatment groups was loaded from biological triplicate cultures into 10% SDS–polyacrylamide gels for the separation of target proteins.

Article Snippet: The BNL CL2 cell line was used as a model system, which was a generous gift from Associate Prof of Medicine Dimiter Avtanski, Zucker School of Medicine at Hofstra/Northwell, Hempstead, New York.

Techniques: Western Blot, Comparison, Expressing, Cell Culture, Multiplex sample analysis, Control

Journal: bioRxiv

Article Title: L-Arginine supplementation modulates L-Arg/NO metabolic processes and AMPK/ACC-1 signalling in BNL CL2 hepatocytes

doi: 10.64898/2026.02.03.703662

Figure Lengend Snippet: Relative total and phosphorylated protein expressions of AMPK and ACC-1 in BNL CL2 cells with exogenous L-Arg Relative protein amounts for total AMPKα (a) and ACC-1 (b) , phosphorylated AMPKα at Thr172 (AMPKα-P) (c) and ACC-1 at Ser79 (AMPKα-P) (d) and the ratio of AMPKα-P to total AMPKα in BNL CL2 cells (e) and ACC-1-P to total ACC-1 in BNL CL2 cells (f) . Cells were cultured for 24 or 72 h in customized media containing 0, 400 and 800 µM L-Arg. Controls were addition of complete DMEM or untreated cultures at T=0. Bands in the blots were quantified using ImageJ software and the data are normalised to β-actin and then expressed as relative to the no L-Arg SILAC DMEM (0 µM L-Arg) control at 24 h. Data points represent the mean ± SD. Error bars represent the standard deviation from the mean (n = 3).

Article Snippet: The BNL CL2 cell line was used as a model system, which was a generous gift from Associate Prof of Medicine Dimiter Avtanski, Zucker School of Medicine at Hofstra/Northwell, Hempstead, New York.

Techniques: Cell Culture, Software, Multiplex sample analysis, Control, Standard Deviation

Journal: bioRxiv

Article Title: L-Arginine supplementation modulates L-Arg/NO metabolic processes and AMPK/ACC-1 signalling in BNL CL2 hepatocytes

doi: 10.64898/2026.02.03.703662

Figure Lengend Snippet: Western blot comparison of the expression of key protein; AMPK and lipogenic ACC-1 protein levels investigated in L-Arg/NO metabolic pathway signalling in BNL CL2 cells cultured in customized media containing (a) no L-Arg SILAC DMEM and L-NAME (4 mM), (b) complete DMEM and L-NAME (4 mM), (c) 400 µM L-Arg and L-NAME (4 mM), and (d) 800 µM L-Arg and L-NAME (4 mM) for 24 and 72 h. Relative protein expression of AMPK (e) and ACC-1 (f) involved in L-Arg/NO metabolic pathway signalling in BNL CL2 cells cultured in different concentrations of L-Arg and the control with or without L-NAME for 24 and 72 h conditions investigated. β-actin is used as a loading control. The same amount of protein (10 µg) from different treatment groups was loaded from biological triplicate cultures into 10% SDS–polyacrylamide gels for the separation of target proteins. Data points represent the mean ± SD. Error bars represent the standard deviation from the mean (n = 3).

Article Snippet: The BNL CL2 cell line was used as a model system, which was a generous gift from Associate Prof of Medicine Dimiter Avtanski, Zucker School of Medicine at Hofstra/Northwell, Hempstead, New York.

Techniques: Western Blot, Comparison, Expressing, Cell Culture, Multiplex sample analysis, Control, Standard Deviation

Journal: bioRxiv

Article Title: L-Arginine supplementation modulates L-Arg/NO metabolic processes and AMPK/ACC-1 signalling in BNL CL2 hepatocytes

doi: 10.64898/2026.02.03.703662

Figure Lengend Snippet: Western blot comparison of the expression of key protein; AMPK and lipogenic ACC-1 protein levels investigated in L-Arg/NO metabolic pathway signalling in BNL CL2 cells cultured in customized media containing (a) no L-Arg SILAC DMEM and SNAP (100 µM) (b) complete DMEM and SNAP (100 µM), (c) 400 µM L-Arg and SNAP (100 µM), and (d) 800 µM L-Arg and SNAP (100 µM) for 6 and 24 h. Relative protein expression of AMPK (e) and ACC-1 (f) involved in L-Arg/NO metabolic pathway signalling in BNL CL2 cells cultured in different concentrations of L-Arg and the control with or without SNAP for 24 h conditions investigated. β-actin is used as a loading control. The same amount of protein (10 µg) from different treatment groups was loaded from biological triplicate cultures into 10% SDS–polyacrylamide gels for the separation of target proteins.

Article Snippet: The BNL CL2 cell line was used as a model system, which was a generous gift from Associate Prof of Medicine Dimiter Avtanski, Zucker School of Medicine at Hofstra/Northwell, Hempstead, New York.

Techniques: Western Blot, Comparison, Expressing, Cell Culture, Multiplex sample analysis, Control

Journal: bioRxiv

Article Title: L-Arginine supplementation modulates L-Arg/NO metabolic processes and AMPK/ACC-1 signalling in BNL CL2 hepatocytes

doi: 10.64898/2026.02.03.703662

Figure Lengend Snippet: Cell culture supernatant was obtained from cultured BNL CL2 cells grown in the presence of 0, 400 or 800 µM L-Arg and the controls complete DMEM addition for 24 or 72 h and untreated cell samples at T=0 (relative (a) and actual (b) values), cells grown in the presence of L-NAME (4 mM, relative (c) and actual (d) values) and cells grown in the presence of SNAP (100 µM) in 0, 400 or 800 µM L-Arg and the control complete DMEM for 6 or 24 h (relative (e) and actual (f) values). Quantified nitrite was normalised to the nitrite amount presence in cultures with no L-Arg SILAC DMEM (0 µM L-Arg) at 24 h. Data points represent the mean ± SD. Error bars represent the standard deviation from the mean (n = 3).

Article Snippet: The BNL CL2 cell line was used as a model system, which was a generous gift from Associate Prof of Medicine Dimiter Avtanski, Zucker School of Medicine at Hofstra/Northwell, Hempstead, New York.

Techniques: Cell Culture, Control, Multiplex sample analysis, Standard Deviation

Journal: bioRxiv

Article Title: L-Arginine supplementation modulates L-Arg/NO metabolic processes and AMPK/ACC-1 signalling in BNL CL2 hepatocytes

doi: 10.64898/2026.02.03.703662

Figure Lengend Snippet: Relative and actual residue amount of L-Arg (a and b) , L-Cit (c and d) and (e and f) analysed in plasma samples of BNL CL2 cells cultured in different amount of L-Arg (0, 400 and 800 µM) and the control complete DMEM media after 24 and 72 h. Untreated cultures at T=0. Data points represent the mean ± SD of each sample. Error bars represent the standard deviation from the mean (n =3).

Article Snippet: The BNL CL2 cell line was used as a model system, which was a generous gift from Associate Prof of Medicine Dimiter Avtanski, Zucker School of Medicine at Hofstra/Northwell, Hempstead, New York.

Techniques: Residue, Clinical Proteomics, Cell Culture, Control, Standard Deviation

Journal: bioRxiv

Article Title: L-Arginine supplementation modulates L-Arg/NO metabolic processes and AMPK/ACC-1 signalling in BNL CL2 hepatocytes

doi: 10.64898/2026.02.03.703662

Figure Lengend Snippet: Relative and actual residue amount of L-Arg (a and b) , L-Cit (c and d) and L-Orn (e and f) analysed in plasma samples of BNL CL2 cells cultured with or without the presence of iNOS inhibitor; L-NAME (4 mM) in excess amount of L-Arg (400 and 800 µM) after 24 and 72 h. Data points represent the mean ± SD of each sample. Error bars represent the standard deviation from the mean (n =3).

Article Snippet: The BNL CL2 cell line was used as a model system, which was a generous gift from Associate Prof of Medicine Dimiter Avtanski, Zucker School of Medicine at Hofstra/Northwell, Hempstead, New York.

Techniques: Residue, Clinical Proteomics, Cell Culture, Standard Deviation

Journal: bioRxiv

Article Title: L-Arginine supplementation modulates L-Arg/NO metabolic processes and AMPK/ACC-1 signalling in BNL CL2 hepatocytes

doi: 10.64898/2026.02.03.703662

Figure Lengend Snippet: Relative and actual residue amount of L-Arg (a and b) , L-Cit (c and d) and L-Orn (e and f) analysed in plasma samples of BNL CL2 cells cultured with or without the presence of external NO donor; SNAP (100 µM) in excess amount of L-Arg (400 and 800 µM) after 6 and 24 h. Data points represent the mean ± SD of each sample. Error bars represent the standard deviation from the mean (n =3).

Article Snippet: The BNL CL2 cell line was used as a model system, which was a generous gift from Associate Prof of Medicine Dimiter Avtanski, Zucker School of Medicine at Hofstra/Northwell, Hempstead, New York.

Techniques: Residue, Clinical Proteomics, Cell Culture, Standard Deviation

Journal: bioRxiv

Article Title: L-Arginine supplementation modulates L-Arg/NO metabolic processes and AMPK/ACC-1 signalling in BNL CL2 hepatocytes

doi: 10.64898/2026.02.03.703662

Figure Lengend Snippet: The proposed mechanisms responsible for the beneficial effect of L-Arg/NOS/NO on the metabolic pathways in mammalian liver cells, BNL CL2. The symbol (+) denotes an upregulation in gene expression or protein expression. The symbol (-) denotes a downregulation of gene expression or protein expression.

Article Snippet: The BNL CL2 cell line was used as a model system, which was a generous gift from Associate Prof of Medicine Dimiter Avtanski, Zucker School of Medicine at Hofstra/Northwell, Hempstead, New York.

Techniques: Gene Expression, Expressing