Fig. 5

Fig. 5

PTBP1 decoy oligonucleotides inhibit its splicing and biological activities in breast cancer cells. a Western blot of proteins pulled down with biotin conjugated PTBP1i (biPTBP1i) or SCRM (biSCRM) using HEK293 nuclear extracts. b Volcano plot showing statistically significant proteins pulled down with biotin conjugated PTBP1i, compared to SCRM control, using HEK293 nuclear extracts. c RT-PCR and quantification of known PTBP1 splicing targets in MDA-MB-231 cells 48 h after transfection with either 5 µM biSCRM or biPTBP1 oligonucleotides. **p-value <0.0008. Data represent means ± SD of five independent biological samples. d Proliferation assay of MDA-MB-231 cells transfected as in c. Four hours after transfection 4000 cells/well were seeded in six replicates. **p-value <0.0002 for all time points. Data represent means ± SD of six replicates. e Clonogenic assay of MDA-MB-231 cells transfected as in c plated at different densities (250, 500, or 1000 cells/well). n = 2. Data represent means ± SD of duplicate wells. f Soft agar colony growth assay on MDA-MB-231 cells transfected as in c. Graphs represent quantification of 10 fields counted in duplicate (total of 20 fields). **p-value <1.7E−27. Data represent means ± SD of two wells. p-values were calculated using Student's t-test (two-tailed)