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. Author manuscript; available in PMC: 2011 Jun 28.
Published in final edited form as: Mol Cell. 2010 Mar 12;37(5):620–632. doi: 10.1016/j.molcel.2010.02.018

Figure 6. Glucose and LKB1 Regulate miR-451 Levels, and miR-451 Reduction Is Critical for the Cellular Response to Glucose Withdrawal.

Figure 6

(A) Effect of glucose deprivation on miR-451 expression levels. Cells were cultured for 24 hr in normal (+, glucose 4.5 g/l) or reduced glucose (−, glucose 0.3 g/l) medium. miR-451 levels were then measured by qRT-PCR and expressed as mean relative to normal medium controls ± SD.

(B) Effect of AMPK and LKB1 downregulation on miR-451 expression. U87, U251,and U373 glioma cells were transfected with negative control siRNA, AMPKα1 siRNA, or LKB1 siRNA. miR-451 levels were measured 48 hr after transfection by qRT-PCR and expressed as mean relative to controls ± SD (left). Corresponding western blots with anti-AMPK, anti-LKB1, and anti-α-tubulin antibody as a loading control are shown below (right).

(C) Effect of miR-451 expression on glucose deprivation-dependent AMPK activation. U251 cells were transfected either with negative control miR (NC) or miR-451 precursor and grown in normal (+, glucose 4.5 g/l) or reduced glucose (−, glucose 0.3 g/l) medium for 18 hr. Cell lysates were blotted with anti-CAB39, anti-phospho-AMPK, anti-AMPK, anti-phospho-Raptor, and anti-Raptor antibodies. Anti-α-tubulin antibody was used as a loading control.

(D) Effect of miR-451 expression on cellular response to glucose deprivation. U251 cells were transfected either with negative control miR (NC) or miR-451 precursor and were grown in normal (+, glucose 4.5 g/l) or reduced glucose (−, glucose 0.3 g/l) medium for 24 hr. Cell number was quantified by crystal violet staining and was expressed as mean cell number relative to controls ± SD. *p < 0.01. See also Figure S3.