MDA-MB-231 cells grown in six-well plates at 37 °C and treated with the tribodies at a concentration of 200 nM for 72 h were collected and lysed, as previously reported [44 (link)]. Protein concentrations were determined by Bradford colorimetric assay (Biorad, California, USA), and Western blotting analyses were performed by incubating the membranes with the anti-phospho-p44/42 MAPK, anti-p44/42 MAPK, anti-phospho-Sapk/Jnk, anti-Sapk/Jnk, anti-cleaved Caspase-3, anti-Caspase 3 and anti-vinculin antibodies (Cell Signaling Technology, Danvers, MA, USA), followed by incubation with the appropriate HRP-conjugated secondary antibodies and detection, as previously reported [45 (link)].
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