Triptolide and C646 were purchased from Selleck chemicals. HSF1 expression plasmid (HSF1-GFPN3) was obtained from Addgene. Mec-1 cells were purchased from DSMZ. Mec-2 and WAC3-CD5+ cells were obtained from Dr. H. Shi (GRU Cancer Center, GA). All cells were cultured as described previously and authenticated by surface staining and flow cytometry for CD19, CD20 and CD79a [53 (link)]. Stable knockdown of HSF1 was carried out in Mec-1 and WaC3-CD5+ cells using lentiviral non-targeted or two independent HSF1 shRNA constructs (Sigma Aldrich, MO). De-identified and de-linked primary CLL samples were obtained from the Biorepository Core Facility of Kansas University Medical Center, after informed consent using an institutional review board-approved protocol (HSC-5929). CD19+ B cells from newly diagnosed, relapsed or treatment refractory CLL patients were isolated from the samples utilizing a magnetic CD19-positive selection kit (Stem Cell Technologies, Vancouver, BC), as described previously [54 (link)]. The purity of the isolated CD19+ B-cell fraction was assessed using CD19-PE conjugated antibodies (BD Biosciences, San Jose, CA) and flow cytometry. Positively selected cells were re-suspended in 20% FBS containing RPMI prior to performing the studies described.
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