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. Author manuscript; available in PMC: 2011 Jun 15.
Published in final edited form as: Dev Cell. 2010 Jun 15;18(6):1041–1052. doi: 10.1016/j.devcel.2010.05.005

Figure 6.

Figure 6

Regulation of autophagy by cytokines is dependent on the type III PI3 kinase. A–B, Quantification of PtdIns3P levels in H4 FYVE-dsRed cells grown in serum-free medium supplemented for 24h with 100 ng/mL IGF1, 50 ng/mL FGF2, 50 ng/mL LIF, 50 ng/mL CLCF1 or 50 ng/mL SDF1, in the absence (A) or presence (B) of 50 nM rapamycin. C, Quantification of PtdIns3P levels in H4 FYVE-dsRed cells treated with indicated cytokines in the presence of the type III PI3 kinase inhibitor 3MA (10 mM). D, Quantification of levels of autophagy following cytokine treatment of Beclin 1 knock-down H4 LC3-GFP cells. E–F, Induction of PtdIns3P levels in H4 FYVE-dsRed cells (E) and up-regulation of autophagy in H4 cells (F) following siRNA mediated knock-down of indicated genes is attenuated in the presence of 3MA. Cells were transfected with indicated siRNAs for 72h, 10 mM 3MA was added for 8h before cells were processed for analysis. For western blots 10 μg/mL E64d was added and quantification of LC3 II/tubulin ratio is shown. **p<0.01, n≥6 All error bars are s.e.m. G, Model for opposing regulation of cell growth and autophagy by the type I and type III PI3 kinases in response to changes in extracellular environment.