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. 2016 May 5;7(23):34785–34799. doi: 10.18632/oncotarget.9185

Figure 4. LY2603618 treatment results in decreased expression of Mcl-1.

Figure 4

(A) CTS and U937 cells were treated with LY for 24 h. Whole cell lysates were subjected to Western blotting. (B and C) Relative densitometry of Mcl-1 expression were measured using Odyssey Software V3.0 and graphed as fold change compared to the no drug control ± s.e.m. *indicates p < 0.05, **indicates p < 0.005, and ***indicates p < 0.0005. (D) CTS and U937 cells were treated with LY and ROSC alone or in combination for 24 h. Whole cell lysates were subjected to Western blotting. The fold changes for the Mcl-1 densitometry measurements, normalized to β-actin and then compared to no drug treatment control, are indicated. (E) U937 cells were infected with Precision LentiORF Mcl-1 (U937/Mcl-1) and RFP control (U937/RFP) lentivirus particles overnight, then washed and incubated for 48 h prior to adding puromycin to the culture medium. The puromycin-resistant cells were treated with LY for 24 h. Whole cell lysates were subjected to Western blotting. The fold changes for the Mcl-1 densitometry measurements, normalized to β-actin and then compared to no drug treatment control, are indicated. (F) U937/RFP and U937/Mcl-1 cells were treated with LY for 24 h and then subjected to Annexin V/PI staining and flow cytometry analysis. **indicates p < 0.005 and ***indicates p < 0.0005. (G) U937 cells were treated with 4 μM LY for the indicated times. Whole cell lysates were subjected to Western blotting. Mcl-1 densitometry measurements, normalized to β-actin and then compared to no drug treatment control at the corresponding time point, are indicated. (H) U937 cells were treated with LY for 2 h, 4 h, and 8 h and then subjected to Annexin V/PI staining and flow cytometry analysis. **indicates p < 0.005 and ***indicates p < 0.0005. (I) U937 cells were treated with LY and ROSC, alone or in combination, for 8 h. Cells were then subjected to Annexin V/PI staining and flow cytometry analysis. ***indicates p < 0.0005. (J) Whole cell lysates were subjected to Western blotting. Densitometry measurements, normalized to β-actin and then compared to no drug treatment control, are indicated.