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. 2018 Nov 19;9(12):1147. doi: 10.1038/s41419-018-1177-6

Fig. 6. CCND1 and BCL2, both are essential for TFF3-driven oncogenic transformation of immortalized-HMECs.

Fig. 6

a Western blot analysis was used to assess the levels of CCND1 and BCL2 in immortalized-HMECs with forced expression of TFF3 and their vector control cells after siRNA-mediated depletion of CCND1 or BCL2. A control cell was transiently transfected with scrambled oligo. Soluble whole cellular extracts were run on an SDS-PAGE and immunoblotted as described in materials and methods. β-actin was used as an input control for cell lysate. The sizes of detected protein bands in kDa are shown on the right side. b Cell viability of immortalized-HMECs with forced expression of TFF3 and their vector control cells in 3D Matrigel culture after depletion/inhibition of CCND1 and BCL2. Depletion of CCND1/BCL2 was executed using CCND1-siRNA/ BCL2-siRNA, respectively. Inhibition of CCND1 or BCL2 was executed upon exposure to Arcyriaflavin A (AA) (50 nM) or YC137 (1 µM) inhibitor, respectively. c BrdU incorporation in immortalized-HMECs with forced expression of TFF3 and their vector control after depletion/inhibition of CCND1 and BCL2. Depletion of CCND1/BCL2 was executed using CCND1-siRNA/ BCL2-siRNA, respectively. Inhibition of CCND1 or BCL2 was executed upon exposure to Arcyriaflavin A (AA) (50 nM) or YC137 (1 µM) inhibitor, respectively. d Caspase 3/7 activity in immortalized-HMECs with forced expression of TFF3 and their vector control cells after depletion/inhibition of CCND1 and BCL2. Depletion of CCND1/BCL2 was executed using CCND1-siRNA/ BCL2-siRNA, respectively. Inhibition of CCND1 or BCL2 was executed upon exposure to Arcyriaflavin A (AA) (50 nM) or YC137 (1 µM) inhibitor, respectively. All assays were performed as described in Material and Methods. The column is mean of triplicate experiments; bars, ±SD. **P < 0.001, *P < 0.05