Taxol t7402

Chemical and standard reagents including filipin and NBD-cholesterol were obtained from Sigma-Aldrich, St. Louis, MO. LysoTracker and fluorescent dextran were obtained from Life Technologies (Grand Island, NY). shRNA constructs were obtained from SA Biosciences (Valencia, CA). Antibodies against tubulin [1:100 for immunofluorescence (IF), T6199, Sigma-Aldrich], EGFP (1:100 for IF, 632380, Clontech) and mCherry (1:100 for IF, NBP1-96752, Novus Biologicals) have been described previously (Whyte et al., 2008 (link); Towns et al., 2009 (link)). Antibodies against StARD9 [1:100 for western blotting (WB), HPA014562] were obtained from Sigma-Aldrich. Site-directed mutagenesis utilized the Phusion kit (New England Biolabs, Ipswich, MA). Antibodies against NPC1 (5 μg/ml for IF, 1 μg/ml for WB, ab108921), LAMP1 (1:10 for IF, ab25630), SREBP1 (1:100 for IF, ab28481) and cathepsin B (1 μg/ml for IF, ab6313) were obtained from Abcam (Boston, MA). filipin (f9765) was obtained from Sigma-Aldrich. NBD-cholesterol (N1148) was obtained from Invitrogen (Waltham, MA). BacMAM-ER (C10590) was obtained from Thermo Fisher Scientific (Waltham, MA). Nocodazole (M1404) and taxol (T7402) were obtained from Sigma-Aldrich. The sequences of oligonucleotides used in this study are provided in Table S3.

The cells were treated with recombinant human TGFβ1 (5 ng·mL⁻¹, PeproTech EC Ltd, London, UK) and recombinant human BMP7 (100 ng·mL⁻¹, a gift from K. Sampath, Sanofi‐Gelisa Research Center, Framingham, MA, USA) for the time periods indicated in the Figures. The LY2157299 TβRI kinase inhibitor (Galunisertib; Sigma‐Aldrich AB, Stockholm, Sweden) was administered to cells at a final concentration of 2.5 μm. Anti‐cancer drugs, 5´‐fluoro‐uracil (5‐FU, F6627, Sigma‐Aldrich AB, Stockholm, Sweden), cisplatin (232120, Sigma‐Aldrich AB, Stockholm, Sweden), cyclophosphamide (Toronto Research Chemicals, North York, Canada), doxorubicin (D1515, Sigma‐Aldrich AB, Stockholm, Sweden) and taxol (T7402, Sigma‐Aldrich AB, Stockholm, Sweden) were used for chemoresistance experiments. Dimethyl‐sulfoxide (DMSO) served as a vehicle for all chemicals and additional details are presented in Table S3.