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. 2016 Jun 13;7(30):47687–47698. doi: 10.18632/oncotarget.9959

Figure 5. The LACE1-mediated apoptosis is dependent on p53.

Figure 5

A. Transfection of human dermal fibroblasts with LACE1 stealth siRNA leads to efficient knockdown of LACE1 protein. Normal human dermal fibroblasts (NHDFs) were transfected with control and LACE1 siRNA, harvested 48 hours post-transfection and processed for western blotting. The blots were developed with antibodies to LACE1 and mtHSP70. B. Knockdown of LACE1 in NHDFs by siRNA renders cells resistant towards STS-induced apoptosis. Both control siRNA- and LACE1 siRNA-transfected NHDFs were treated with 0-h, 3-h and 6-h exposures to staurosporine (2 μM). Whole-cell lysates were subsequently western blotted with antibody to cleaved PARP. Equal protein loading was controlled with antibody to α-tubulin. C. The LACE1-mediated apoptosis is dependent on p53. NHDFs in which p53 was downregulated using siRNA as well as appropriate siRNA control NHDFs were transfected with either LACE1-FLAG or empty vector constructs. The resulting whole-cell extracts were western blotted with antibodies to p53, cleaved PARP, FLAG. Equal protein loading was monitored with α-tubulin detection.