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

Figure 4.

Figure 4

HPSE reduces nuclear PTEN level. (A) RPMI-8226 cells were treated with PBS as a control (Cont) or rHPSE (1 µg/mL) for the designated time followed by isolation of cytoplasmic and nuclear fractions. PTEN present in each fraction was assessed by Western blotting. LaminB1 and GAPDH were used as loading control and to assess the quality of the isolated fractions. The bar graphs show the mean relative expression of PTEN normalized with GAPDH for the cytoplasmic fraction or normalized with LaminB1 for the nuclear fraction in three independent experiments. ** p < 0.01 and * p < 0.05; NS—not significant. (B) CAG HPSE Hi cells were treated with OGT2115 and the levels of PTEN and HPSE present in the cytoplasmic and nuclear fractions were evaluated by Western blot. LaminB1 and GAPDH were utilized for assessing the quality of the fractions. (C) HPSE Hi cells were incubated without or with the heparanase inhibitor OGT2115 for 16h. PTEN, CyclinD1 (CCND1) and HPSE expression were evaluated in whole cell lysates (WCL) by Western blot.