Y705 phosphorylation is needed for the correct localisation of STAT3 in the mitochondrion. (A) IF with anti-STAT3 and anti-ATAD3 antibodies on ESCs transiently transfected with either mStat3, mStat3 Y705F or MLS_mStat3_NES Y705F. Arrowheads indicate the colocalisation of ATAD and STAT3. (B) Western blot of total STAT3 in ESC extracts; β-actin was used as a loading control. (C) Western blot of mitochondrial STAT3 from ESC mitochondrial extracts; VDAC1 was used as a mitochondrial loading control, Lamin was used as a control for nuclear contamination. (D) Representative pictures of DAB IHC on ESCs acquired with TEM; positive signal is black and negative is white. Cristae are positive in Stat3 WT and MLS_Stat3_NES transfected cells. m, mitochondria; n, nucleus. Scale bar: 200 μm (A).
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