Extended Data Figure 7: The epichaperome facilitates cancer cell survival.

The expression of the epichaperome was altered by epichaperome components knockdown (a) or by titrating into cells siRNAs that targeted both the HSP90α and the HSP90β paralogues (b–g) to test whether the epichaperome facilitates survival in type 1 tumours. a, Epichaperome levels were altered by AHA1 siRNA knockdown or a control (scramble, Scr) siRNA (left panel) and cell viability, as measured by PARP cleavage, was determined in cells treated for 24 h with increasing concentrations of PU-H71 (0, 0.5, 1 and 2 μM) (right panel). See also Extended Data Fig. 5g for biochemical signature of cells after AHA1 knockdown. Data are representative of two independent experiments. b–e, Total protein (b, e), mRNA (c) and multimeric species of indicated chaperome members (d) levels were monitored in MDA-MB-468 type 1 cells in which several concentrations of siRNAs against HSP90α/β (n = 14) were titrated in. 1 and 8 are control scramble; 2, 3, 4, 9, 10, 11 are 0.915, 1.83, 3.66, 0.366, 1.83 and 14.64 nM of siRNA no. 1, respectively; and 5, 6, 7, 12, 13, 14 are 0.366, 0.915, 3.66, 0.0229, 0.0915 and 0.366nM of siRNA no. 2, respectively. Cell viability in each condition was monitored by LDH release. Values for each experimental condition (as percentage control scramble) were quantified and are noted under the native gels in a. HMW, high molecular weight. For gels, experiments were repeated independently twice with representative data shown. For graphs, mean ± s.d., n = 6. f, Changes in chaperome members (mRNA) were monitored following siRNA knockdown of the indicated individual chaperome members in MDA-MB-468 and ASPC1 cells. Error bars show mean ± s.d., n = 6. g, Same as for b–d in ASPC1, type 2 cells (1 through 4 siRNA concentrations, as in d. Error bars represent mean ± s.d., n = 6, unpaired t-test, NS, not significant). For uncropped gel data, see Supplementary Fig. 1.

Source data