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. 2017 Sep 21;8(56):95038–95053. doi: 10.18632/oncotarget.21119

Figure 2. Ppp4r2 suppression regulates DNA damage response in normal murine hematopoietic cells.

Figure 2

(A) Impact of Ppp4r2 knockdown in murine Lin- bone marrow (BM) cells on clonogenic growth determined by colony forming cells in methylcellulose (CFC-Assay; n = 4). (B) Impact of Ppp4r2 knockdown in murine Lin- BM cells on DNA damage response upon ionizing radiation (IR) with 2 Gy determined by the mean fluorescence intensity (MFI) of phosphorylated KRAB-domain associated protein 1 [pKAP1(S824)] at the indicated time post IR (n = 2). Representative histogram depicts the MFI shift of pKAP1 (S824) at 0.5 h post IR in either murine Lin- BM cells with Ppp4r2 knockdown or control. (C) DNA damage at indicated time post IR with 2 Gy in murine Lin- BM cells upon Ppp4r2 knockdown determined by the MFI of phosphorylated histone variant H2AX [γH2AX (S139); n = 3]. Representative histogram depicts the MFI shift of γH2AX (S139) at 0.5 h post IR in murine Lin- BM cells with either Ppp4r2 knockdown or control. (D) Representative Western Blot displaying the effect of IR on phosphorylation of KAP1 (S824) and P53 (S15) in Lin- BM cells with either Ppp4r2 knockdown or control. Vertical lines have been inserted to indicate a repositioned gel lane. (E) Apoptosis induction upon IR with 2 Gy displayed as the percentage of AnnexinV+/7AAD- Lin- BM cells with either Ppp4r2 knockdown or control at the indicated time post IR (n = 3). Data are represented by the mean ± SD. Statistical analyses were carried out using unpaired two-tailed t-test or multiple t-tests corrected for multiple comparisons using the Holm-Sidak method. A p-value ≤ 0.05 was considered significant, *p ≤ 0.05.