Fig. 4: KRASG12D upregulates YB-1 in de novo KRASG12D-transformed normal human mammary cells.
A Western blots showing YB-1 levels (relative to H3) in human BCs, LPs, LCs and stromal cells (SCs) isolated viably from three normal donors according to their differential surface expression of EPCAM and CD49f levels and absence of expression of CD45 and CD31 (top panel). B Representative views of YB-1 immunostaining of normal human mammary tissue (left) and 8-week tumours derived from KRASG12D-transduced normal mammary cells isolated from the same three normal donors (right). Scale bar, 50 μm. Bar graph shows quantification of YB-1 expression. Data are from individual tumours (Normal, n = 8; de novo tumours, n = 8). C Representative FACS profile of a 4-week xenograft of inducible KRASG12D-transduced human BCs obtained from mice maintained post-transplant on doxycyline-supplemented water (Dox) for just the first 2 weeks (left panel) or for the full 4 weeks (right panel) of the experiment. D KRAS mRNA levels measured in 4-week xenografts of inducible KRASG12D-transduced cells obtained from mice maintained on Dox for the first 2 weeks only, or the full 4 weeks of the experiment, as shown. The dot plot shows KRAS relative expression compared to the No Dox control mice (n = 2), as ΔΔCt values. E Representative views of YB-1 immunostaining of 4-week tumours derived from mice transplanted with cells transduced with a Dox-inducible KRASG12D cDNA and maintained on Dox for the just the first 2 weeks or the full 4 weeks before the tumours were harvested for analysis (N = 3 donors). Scale bar, 50 μm.
