Figure 2: AK023948 regulates the pAKT level.
(a) While AK023948 overexpression increases, AK023948 knockdown decreases phosphorylation of AKT. MCF-7 cells were transfected with either AK023948 expression vector or AK023948 siRNA. Cellular extract was prepared for western blot 48 h after transfection. (b) AK023948 KO suppresses the pAKT level. AK023948 KO was performed in MCF-7 using a procedure as described in Methods. (c) Re-expression of AK023948 in the AK023948 KO cells increases the pAKT level. AK023948 expression vector or control vector was introduced into KO cells, and total cellular extract was prepared for western blot. (d) High levels of AK023948 and pAKT in breast tumour specimens with representative images for the same field of a tumour. The same TMA was first detected for AK023948 by ISH and then the signal was stripped, followed by IHC to detect pAKT. Scale bar, 100 μm. (e) A positive correlation between AK023948 and pAKT from the same TMA with P-value of Fisher’s exact test for the association <0.0001. (f) The pAKT level is lower in xenograft tumours derived from AK023948 KO than in vector control tumours, as detected by IHC. Scale bar, 50 μm.
