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. 2017 Jun 13;9:429–436. doi: 10.2147/BCTT.S135665

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© 2017 Hou et al. This work is published and licensed by Dove Medical Press Limited

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High glucose regulates EGFR activity through GTPase.

Notes: (A) Serum-deprived MDAMB231 cells expressing scrambled (lanes 1 and 2) or Rac1 RNAis (lanes 3 and 4) were cultured in low glucose condition (5 mM) for 12 hours and then exposed to high glucose (25 mM). After 6 hours, these cells were harvested for the measurement of EGFR phosphorylation, total EGFR levels and EGFR ubiquitination. The Western blot analysis of EGFR phosphorylation was quantified and presented by the plot at the bottom. (B) Serum-deprived MDAMB231 cells expressing scrambled (lanes 1 and 2) or Cdc42 RNAis (lanes 3 and 4) were cultured in low glucose condition (5 mM) for 12 hours and then exposed to high glucose (25 mM) with serum (5% FBS). After 6 hours, these cells were harvested for measurement of EGFR phosphorylation, total EGFR levels, the ubiquitination of EGFR, c-cbl phosphorylation, and total c-cbl levels. The Western blot analysis of EGFR phosphorylation and EGFR ubiquitination were quantified and presented by the plots at the bottom. (C) Serum-deprived MDAMB231 cells expressing HA-tagged Rac1 (G12V) (lane 2) were cultured in low glucose condition (5 mM) for 12 hours and then were harvested for measurement of EGFR phosphorylation and total EGFR levels. The Western blot analysis of EGFR phosphorylation was quantified and presented by the plot at the bottom. (D) Serum-deprived MDAMB231 cells expressing HA-tagged Cdc42 (F28L) (lane 2) were cultured in low glucose condition (5 mM) for 12 hours and then exposed to high glucose (25 mM, lane 3) with serum (5% FBS). After 6 hours, these cells were harvested for measurement of EGFR phosphorylation, total EGFR levels, and the ubiquitination of EGFR. The Western blot analysis of EGFR phosphorylation and EGFR ubiquitination were quantified and presented by the plots at the bottom.

Abbreviations: EGFR, epidermal growth factor receptor; FBS, fetal bovine serum; HG, high glucose; LG, low glucose; WB, western blot; WCL, whole cell lysates.