Fig 2: CDC37 is responsible for reactivation of Akt in mutant BRAF colon cancer cells after treatment with AUY922(A) Whole cell lysates from Lim1215, Caco-2, RKO and WiDr treated with AUY922 (400 nM) for indicated time points were subjected to Western blot analysis. Data are representative, n = 3. (B) Whole cell lysates from RKO (left) and WiDr (right) cells transfected with the control or CDC37 siRNAs were treated with AUY922 (400 nM) for 48 hours and subjected to Western blot analysis. Data are representative, n = 3. (C) Left panel: Whole cell lysates from WiDr cells treated with AUY922 (400 nM) for 24 hours were subjected to immunoprecipitation with a mouse antibody against CDC37. The resulting precipitates were subjected to Western blot analysis. Right panel: Whole cell lysates from WiDr cells transfected with the control or CDC37 siRNA1 were subjected to immunoprecipitation with a rabbit antibody against HSP90. The resulting precipitates were subjected to western blot analysis. Data are representative, n = 3. (D) RKO (left) and WiDr (right) cells transfected with the control or CDC37 siRNAs were treated with AUY922 (400 nM) for 48 hours. Apoptosis was measured by PI and Annexin V staining. Data are mean ± SE, n = 3. **P < 0.01, Student's t-test. (E) Whole cell lysates from RKO (left) and WiDr (right) cells transfected with the control or CDC37 siRNAs were treated with AUY922 (400 nM) for 48 hours and subjected to Western blot analysis. Data are representative, n = 3.

Fig 3: Interaction between Hsp90 and its co-chaperone Cdc37 in human sperm. a. Hsp90 was detected in sperm lysates using western blotting and Co-IP with non-immune mouse IgG or antibodies against Cdc37. No interaction was observed between non-immune mouse IgG and Hsp90, whereas a clear interaction was evident between Hsp90 and Cdc37. 17-AAG disturbed the formation of the Hsp90-Cdc37 complex. Results are representative of three independent experiments using samples from three different individuals. b. The relative association of Hsp90 and Cdc37 with or without 5 µM 17-AAG during human sperm capacitation

Fig 4: Interaction of Hsp103 with cochaperone proteins.Purified Hsp90a, Hsp103, Hop, Aha 1 and Cdc37 were mixed as indicated and fractionated by gel filtration chromatography on a Superdex 200 column. Fractions were analyzed by SDS-PAGE. Marker proteins are shown on top (thyroglobulin, 669 kDa; BSA, 67 kDa). The shift of the cochaperones Hop, Aha1 and Cdc37 indicating complex formation with Hsp90a and Hsp103 is marked by red, blue or green dashed lines, respectively. For the Hsp103+Aha1 and Hsp103+Cdc37 interaction runs the cochaperones were blotted with specific antibodies to prove the identity of the shifted Aha1 and Cdc37 bands that indicate complex formation with Hsp103.

Fig 5: CUL5, CDC37, and HSP90AB1 expression in breast cancer(A) CUL5, CDC37, and HSP90AB1 mRNA expression levels in invasive breast carcinoma patients and normal human breast tissues (CUL5, p < 2.2E-16; CDC37, p = 0.0068; HSP90AB1, p < 2.2E-16). (B) Overall survival in patients with CUL5 (p = 1.74E-7), CDC37 (p = 0.642), and HSP90AB1 (p = 0.224) mutation and wild type. (C) CUL5, CDC37, and HSP90AB1 gene alteration in early and metastasis breast cancer patient. (D) CUL5 (p = 0.0198), CDC37 (p = 0.6864), and HSP90AB1 (p = 0.8839) expression and survival in HER2-E/HER2pos (HER2-enriched) subtype breast cancer patients (cutoff at 40th percentile).