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. 2020 Sep 6;9(9):2038. doi: 10.3390/cells9092038

Figure 5.

Figure 5

Heparanase increases PTEN phosphorylation through CK2 and promotes PTEN protein stability and inactivity. (A) RPMI-8226 control cells were treated with 1 ug/mL of recombinant heparanase (rHPSE) for the indicated time and pPTEN, PTEN, p-AKT, AKT and CK2b protein expression were investigated by Western blot. (B) Quantification of Western blot shown in panel A corresponding to the relative expression of pPTEN, PTEN and CK2b normalized with GAPDH expression, from three independent experiments. * p < 0.05 by one tailed Mann Whitney test. (C) RPMI-Mel-R and RPMI-Dox-R were transfected with siRNA control (siCont) or with two non-overlapping siRNAs to silence HPSE and PTEN level evaluated by Western blot. Right panel, HPSE silencing was evaluated by RT-PCR. (D) CK2b levels present in HPSE-high and HPSE-low patient groups. ** p < 0.01 by one-tailed, unpaired t-test. (E) CAG HPSE Hi and melphalan-resistant (Mel-R) and doxycycline-resistant (Dox-R) RMPI-8226 cell lines were incubated with CK2 inhibitor TBB for 6 h and Western blots performed to assess levels of pPTEN, PTEN and CK2b.