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. 2015 Jan 14;6(8):6105–6122. doi: 10.18632/oncotarget.3368

Figure 4. MCL1 protects cells from undergoing apoptosis under steroid-deprived conditions.

Figure 4

(A) Knockdown of MCL1 in LNCaP cells leads to increased apoptosis under steroid-deprived conditions. LNCaP cells were stably transduced with doxycycline-regulated short hairpin (sh) constructs targeting MCL1 (LNCaPdoxshMCL1) or Luciferase (control, LNCaPdoxshLuc). Both cell lines were grown for 72h under full medium conditions (10% FCS) or under steroid-deprived conditions (10% CSS) and expression of sh constructs was induced by addition of 1 μg/ml doxycycline, as depicted. (B) Overexpression of MCL1 rescues DuCaP and VCaP cells partially from undergoing apoptosis when grown in steroid-deprived conditions. DuCaP and VCaP were stably transduced with a doxycycline-regulated overexpression plasmid for MCL1 generating DuCaPdoxMCL1 and VCaPdoxMCL1. Both cell lines were grown for 40h under full medium conditions (10% FCS) or under steroid-deprived conditions (10% CSS) and overexpression of MCL1 was induced by addition of 1 μg/ml doxycycline. (A, B) Samples were analyzed by immunoblotting using antibodies against MCL1, cleaved PARP (cPARP, as an indicator for apoptosis) and GAPDH. Representative immunoblots are shown. Numbers indicate relative ratios of densitometrical analysis of MCL1 or cPARP divided by the reference protein GAPDH, respectively.