Figure 3
From: Morphotype-specific effector functions of Cryptococcus neoformans PUM1
Pum1 post-transcriptionally regulates its own mRNA in a morphotype specific manner.
(a) Northern analysis of PUM1 in the mycPUM1 and mycPUM1-EM strains grown to mid-log phase in YPD. (b) Mid-log-phase cultures of mycPUM1 and mycPUM1-EM were treated with the transcriptional inhibitor 1,10-phenanthroline and allowed to grow at 30 °C. Aliquots of cells were harvested at 15 min intervals for RNA extraction and subjected to Northern blotting for the detection of PUM1 transcript. Log 2 plot of relative expression of PUM1 transcript normalized to the rRNA was used to calculate the mRNA half-life. The graph shown is the representative of three biological replicates. (c) Western analysis for mycPum1 in the mycPUM1 and mycPUM1-EM strains grown to mid-log phase in YPD. The membrane was probed with primary mouse anti-myc antibody followed by secondary IR680-conjugated Donkey anti-mouse antibody. (d) Fluorescence microscopy of mCh-Pum1 from bilateral mating of mCherry-tagged Pum1 complement strains.
