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. 2015 Nov 28;7(2):1675–1686. doi: 10.18632/oncotarget.6423

Figure 4. The use of different siRNAs targeting the same exon 8 region reveals that Hipk2-FL depletion is not sufficient to prompt apoptosis.

Figure 4

(A) Four different siRNAs homologous to a short region of Hipk2 exon 8 present only in the FL isoform were used for target validation. The sequence of each e8-siRNA is highlighted in red. (B) Two CSC lines were transfected with the indicated siRNAs. All siRNAs were used as Stealth RNAi from Invitrogen, with the exception of e8-siRNA#1U, which is a standard unmodified version of e8-siRNA#1. Cells were collected after 72 hrs, counted to assess viability and analyzed by both RT-PCR and WB. Cleaved CASPASE 3 (clvd CASP3) and β-CATENIN (clvd β-CAT) and PARP1 (Clvd PARP) were used as apoptotic markers. (C) Light microscopy pictures of transfected cells taken 96 hrs post-transfection reveal a clearly different phenotypic aspect in cells treated with different siRNA targeting the same RNA region. The percentage of viable cells is reported at the right bottom of each panel. Bars, 100 μm.