Inhibition of HSP90 in B-cell Malignancies

A high-affinity HSP90-inhibitor, AUY922 [30 (link)] was purchased from Selleckchem. Antibodies to HSP90_αβ, PLCγ2, BCAP, CD19, AXL, BCL2, GAPDH and actin were purchased from Santa Cruz Biotechnologies. Antibodies to CD79a, CD79b, LYN, SYK, BTK, AKT, P-ERK1/2, ERK1/2, STAT3, PTPN22, FGFR3, and MCL-1 were purchased from Cell Signaling Technologies. XIAP antibody, chromogen-conjugated antibodies to human CD5 and CD19 or fluorescein isothiocyanate (FITC)-conjugated annexin V were obtained from BD Biosciences or Invitrogen, respectively. Propidium iodide (PI) and other chemicals were purchased from Sigma or Bio-Rad. Replication-deficient lentiviral constructs expressing HSP90-specific shRNA or GFP tagged control scrambled shRNA were purchased from Santa Cruz Biotechnologies.

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Mahmud H., Mendez M., Mukhopadhyay B., Holter-Chakrabarty J, & Ghosh A.K. (2020). HSP90 overexpression potentiates the B-cell receptor and fibroblast growth factor receptor survival signals in chronic lymphocytic leukemia cells. Oncotarget, 11(22), 2037-2046.

Publication 2020

AUY922 BCL2 GAPDH actin P-ERK1/2 ERK1/2 STAT3 PTPN22 FGFR3 MCL-1 XIAP antibody fluorescein isothiocyanate (FITC)-conjugated annexin V Propidium iodide (PI)

Actin Annexin v Antibodies Antibody Auy922 Bcl2 Cd79a Cd79b Chromogen Erk1 Fgfr3 Fluorescein Gapdh Hsp90 Human Isothiocyanate Plcγ2 Propidium iodide Ptpn22 Replication Shrna Stat3

Corresponding Organization : University of Oklahoma Health Sciences Center

Top 5 similar protocols

Variable analysis

independent variables

Knockdown of HSP90 using shRNA

dependent variables

Levels of HSP90, PLCγ2, BCAP, CD19, AXL, BCL2, XIAP, CD79a, CD79b, LYN, SYK, BTK, AKT, P-ERK1/2, ERK1/2, STAT3, PTPN22, FGFR3, and MCL-1

control variables

Concentration of AUY922 (a high-affinity HSP90 inhibitor)
Antibodies used for Western blot analysis
Propidium iodide (PI) and other chemicals
Fluorescein isothiocyanate (FITC)-conjugated annexin V

positive controls

Scrambled shRNA control

negative controls

Untreated cells

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