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Protein abundance of <t>IRS1,</t> IRS2 and IR in HepG2, AML12 and THLE-2 cells. Cells were grown in their respective growth medium (con) or cultured for 24/48 h without additional insulin. ( a ) 15 µg protein was separated and checked by western blot for the presence of IRS1, IRS2 and IR. GAPDH was used as loading control. ( b – d ) Band intensities were normalized for GAPDH ( n = 3; mean ± s.d.; * p < 0.05, con versus 24/48 h Ø Ins in AML12 cells; § p < 0.05, con HepG2 versus con THLE-2; ¶ p < 0.05, con AML12 versus con THLE-2; ¥ p < 0.05, con HepG2 versus con AML12).
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Protein abundance of <t>IRS1,</t> IRS2 and IR in HepG2, AML12 and THLE-2 cells. Cells were grown in their respective growth medium (con) or cultured for 24/48 h without additional insulin. ( a ) 15 µg protein was separated and checked by western blot for the presence of IRS1, IRS2 and IR. GAPDH was used as loading control. ( b – d ) Band intensities were normalized for GAPDH ( n = 3; mean ± s.d.; * p < 0.05, con versus 24/48 h Ø Ins in AML12 cells; § p < 0.05, con HepG2 versus con THLE-2; ¶ p < 0.05, con AML12 versus con THLE-2; ¥ p < 0.05, con HepG2 versus con AML12).
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Protein abundance of IRS1, IRS2 and IR in HepG2, AML12 and THLE-2 cells. Cells were grown in their respective growth medium (con) or cultured for 24/48 h without additional insulin. ( a ) 15 µg protein was separated and checked by western blot for the presence of IRS1, IRS2 and IR. GAPDH was used as loading control. ( b – d ) Band intensities were normalized for GAPDH ( n = 3; mean ± s.d.; * p < 0.05, con versus 24/48 h Ø Ins in AML12 cells; § p < 0.05, con HepG2 versus con THLE-2; ¶ p < 0.05, con AML12 versus con THLE-2; ¥ p < 0.05, con HepG2 versus con AML12).

Journal: Open Biology

Article Title: Suitability of hepatocyte cell lines HepG2, AML12 and THLE-2 for investigation of insulin signalling and hepatokine gene expression

doi: 10.1098/rsob.180147

Figure Lengend Snippet: Protein abundance of IRS1, IRS2 and IR in HepG2, AML12 and THLE-2 cells. Cells were grown in their respective growth medium (con) or cultured for 24/48 h without additional insulin. ( a ) 15 µg protein was separated and checked by western blot for the presence of IRS1, IRS2 and IR. GAPDH was used as loading control. ( b – d ) Band intensities were normalized for GAPDH ( n = 3; mean ± s.d.; * p < 0.05, con versus 24/48 h Ø Ins in AML12 cells; § p < 0.05, con HepG2 versus con THLE-2; ¶ p < 0.05, con AML12 versus con THLE-2; ¥ p < 0.05, con HepG2 versus con AML12).

Article Snippet: After blocking unspecific binding sites, membranes were incubated overnight with primary antibodies: p-Thr-308, rabbit polyclonal, Cat. 9275, Cell Signaling; p-Ser-473, rabbit polyclonal, Cat. 9271, Cell Signaling; AKT, mouse monoclonal, Cat. 610861, BD Biosciences; GAPDH, mouse monoclonal, Cat. 8245, Abcam; IRS1, rabbit polyclonal, Cat. 06248, Millipore; IRS2, rabbit polyclonal, Cat. 3089, Cell Signaling; and insulin receptor protein chain β, rabbit monoclonal, Cat. 3025, Cell Signaling.

Techniques: Quantitative Proteomics, Cell Culture, Western Blot, Control