FIG 6 .

C. neoformans Atm1 contributes to the maintenance of cytosolic Fe-S cluster homeostasis during Cu stress. (A) Atm1-F Rli1-HA (DTY955) and Gal7-Atm1-F Rli1-HA (DTY957) C. neoformans cells were grown for 88 h in SC-Gluc medium to deplete Atm1 protein levels. During the last 16 h, cells were grown in Fe-poor SC-Gluc medium, and cells were radiolabeled with 10 µCi ⁵⁵Fe for 1.5 h before Cu was added and cells were incubated in the presence of both ⁵⁵Fe and Cu for 1 h (see Fig. 4A). Rli1-HA protein was immunoprecipitated from cell extracts with specific antibodies. The amount of coprecipitated ⁵⁵Fe was quantified by scintillation counting. Data are presented relative to the values obtained for samples not treated with Cu. n = 4 (two-way repeated-measures ANOVA test, P = 0.04). (B) Atm1-F Leu1-HA (DTY959) and Gal7-Atm1-F Leu1-HA (DTY961) C. neoformans cells were experimentally processed and analyzed as described for panel A. n =4 (two-way repeated-measures ANOVA test, P = 0.1).