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. Author manuscript; available in PMC: 2019 Apr 1.

Published in final edited form as: Mol Carcinog. 2018 Jan 25;57(4):536–548. doi: 10.1002/mc.22778

Figure 2. — S3-NTDi downregulates expression of STAT3 target genes. (A) HD-MB03 and DAOY cells were treated with increasing concentrations of S3-NTDi (0, 8, 12 and 16 μM) for 24 h. Cells were stimulated with IL-6/sIL-6Rα (50/25 ng/ml) for 20 mins prior to making whole cell extracts (WCE). Expression of MYC, CCND1 and BCl2 were analyzed by Western immunoblot analysis and GAPDH and β-Actin were used as a loading control. (B) HD-MB03 cells were treated with 10 μM S3-NTDi for overnight or left untreated. Cells were stimulated with IL-6/sIL-6Rα for 20 mins prior to harvest. STAT3 regulated gene expressions were analyzed by qRT-PCR. * represents p<0.001, NT: non-treated control. (C) STAT3 activity is knockdown by 75 nM siRNA (SMARTpool) in HD-MB03 cells for 72 h (inset figure) and STAT3 regulated gene expressions was analyzed by qRT-PCR. * represents p<0.005. (D) Expression of PARP cleavage in HD-MB03 cells, treated with 10 μM of S3-NTDi for 0, 4, 6, 8 and 10 h are shown by Western blot. Intensity of cleaved PARP is shown in bar diagram (bottom). (E) HD-MB03 cells were treated with 0, 8 and 10 μM of S3-NTDi overnight. Total RNA isolated from these cells was subjected to qRT-PCR for CHOP expression. The inset figure shows Western blot of CHOP expression. * represents p<0.001.

Figure 2. — S3-NTDi downregulates expression of STAT3 target genes. (A) HD-MB03 and DAOY cells were treated with increasing concentrations of S3-NTDi (0, 8, 12 and 16 μM) for 24 h. Cells were stimulated with IL-6/sIL-6Rα (50/25 ng/ml) for 20 mins prior to making whole cell extracts (WCE). Expression of MYC, CCND1 and BCl2 were analyzed by Western immunoblot analysis and GAPDH and β-Actin were used as a loading control. (B) HD-MB03 cells were treated with 10 μM S3-NTDi for overnight or left untreated. Cells were stimulated with IL-6/sIL-6Rα for 20 mins prior to harvest. STAT3 regulated gene expressions were analyzed by qRT-PCR. * represents p<0.001, NT: non-treated control. (C) STAT3 activity is knockdown by 75 nM siRNA (SMARTpool) in HD-MB03 cells for 72 h (inset figure) and STAT3 regulated gene expressions was analyzed by qRT-PCR. * represents p<0.005. (D) Expression of PARP cleavage in HD-MB03 cells, treated with 10 μM of S3-NTDi for 0, 4, 6, 8 and 10 h are shown by Western blot. Intensity of cleaved PARP is shown in bar diagram (bottom). (E) HD-MB03 cells were treated with 0, 8 and 10 μM of S3-NTDi overnight. Total RNA isolated from these cells was subjected to qRT-PCR for CHOP expression. The inset figure shows Western blot of CHOP expression. * represents p<0.001.