Figure 3
From: Wee1 inhibition potentiates Wip1-dependent p53-negative tumor cell death during chemotherapy
Wee1 partial inhibition potentializes Wip1-dependent cisplatin-induced cytotoxicity in p53-negative tumor cells but not by inducing a mitotic catastrophe. (a) Flow cytometry assay of H2AX phosphorylation of Saos2 osteosarcoma cells following a treatment with 25 μM of CDDP for 28 h after a reverse transfection of a non-targeting siRNA (line) as a control, or Wee1 siRNA (dots). (b) Cytotoxicity assay in Saos2 osteosarcoma cells after inhibition of Wee1 by the chemical compound MK-1775 used at a low concentration of 75 nM 2 h prior a 25 μM CDDP treatment for 72 h, in absence or presence of Wip1, induced by a 1 μg/ml treatment of doxycycline 24 h prior to the chemotherapeutic treatment. (c) Cytotoxicity assay in Saos2 osteosarcoma cells after inhibition of Wee1 by 75nM MK-1775 2 h prior a 15 μM CDDP treatment for 72 h, with or without Wip1, induced by a 1 μg/ml treatment of doxycycline. (d) Immunofluorescence assay targeting the mitosis-specific marker phospho-histone H3. Cells were treated for 16 h with 100 ng/ml nocodazole to block them in mitosis, with triple combination of 25 μM CDDP, 75nM Wee1 inhibitor MK-1775 and 1 μg/ml doxycycline to induce Wip1 overexpression
