Fig. 8: MOLM-13 and OCI/AML2 were stably transduced with 2 independent shRNA targeting FASN (n = 3).
From: Reducing FASN expression sensitizes acute myeloid leukemia cells to differentiation therapy
A–B FASN knockdown efficiency was validated in MOLM-13 (A) and OCI/AML2 (B) by western blotting. C–H MOLM-13 and OCI/AML2 control and FASN knockdown cells were treated with ATRA for 3 days. C–D CD11b surface marker expression was analyzed as in (A–B). E–F Acridine Orange staining analysis was performed as in 7E-G. G–H Western blot analysis of total protein was extracted and subjected to immunoblotting using anti-FASN, anti-pmTOR(Ser2448), anti- pULK1(Ser757), anti-ULK1, anti-pATG13(Ser318) and anti-ATG13 antibodies. Results shown are from at least two biological duplicates. I FASN localizes to lysosomes in AML with mTOR leading to the phosphorylation of TFEB and its sequestration in the cytoplasm. ATRA treatment resulted in FASN degradation by autophagy and thus TFEB translocation to the nucleus and activation of lysosomal biogenesis.
