Figure 5. Impact of IDH2 mutations on glandular architecture and polarity of non-malignant breast epithelial cells.

(A) Impact of forced expression of IDH2^WT or IDH2^R172S on glandular architecture of MCF10A^P (top) and MCF10A^H1047R (bottom) cells grown in 3D basement membrane cultures (scale bar, 0.1 mm). The volume of the MCF10A^P and MCF10A^H1047R acinar structures expressing IDH2^WT and IDH2^R172S were quantified; *P<0.05; ***P<0.001; ****P<0.0001; error bars represent standard deviation of mean. (B) Immunofluorescent analysis of MCF10A^P and MCF10A^H1047R acinar structures expressing IDH2^WT or IDH2^R172S with antibodies against E-cadherin (red), cis-Golgi marker GM130 (green) and nucleus (DAPI, blue, scale bars, 25μm) (left). Arrows highlight cells with reverse polarity. MCF10A^P and MCF10A^H1047R acinar structures showing either apical or reversed polarity were quantified. MCF10A^H1047R IDH2^R172S acini showed significantly more frequently inverted polarity than MCF10A^H1047R vector control and MCF10A^H1047R IDH2^WT acini (*P<0.05 each; t-test).