Glucose restoration resulted in a rapid glycosylation of the immature VEGFR2 band.

<p>(A) Western blot shows the VEGFR2 expression pattern in the cells exposed to 0 mM glucose media for a period of 24 hrs, followed by glucose restoration for 10, 20, 30 mins and the control cells (cells grown in control regular DMEM (25 mM glucose) without prior hypoglycemic exposure) P: MDA-MB-231P; BO: MDA-MB-231BO and BR: MDA-MB-231BR. The mature VEGFR2 band at 200 kDa was detected after 20 mins of glucose restoration which was more obvious in the MDA-MB-231BR (bottom blot). (B) Western blots showing that glucose restoration for 1 hour (after a period of 24 hrs of hypoglycemia) enhanced the formation of the glycosylated band of VEGFR2 (200 kDa) in all three cell lines MDA-MB-231P (P+glu 1hr), MDA-MB-231BO (BO+glu1h) and MDA-MB-231BR (BR+glu1h). The samples indicated by P25, B25 and BR25 are the samples collected from the cells growing in regular DMEM (25 mM glucose) expressing both mature bands of VEGFR2 (200 kDa and 230 kDa). P0, BO0 and BR0 are the MDA-MB-231P, MDA-MB-231BO and MDA-MB-231BR cells respectively, exposed to hypoglycemia for 24 hrs without glucose restoration. All three cell lines expressed only the unglycosylated band of VEGFR2 (147 kDa) when grown in 0 mM DMEM. (C) Western blots depicting the gradual increase in VEGFR2 band size upon glucose restoration in MDA-MB-231BR cells. Note: β-Actin was used in all the blots as a protein loading control.