Figure 1.

Effects of mg-Proto on the nuclear gene transcription. (a) effects of mg-Proto on the C. merolae nuclear transcriptome. C. merolae culture, after the second 18 hours dark period of the 18 h-dark/6 h-light cycles with air-bubbled conditions, were incubated for 1 hour in the dark in the absence or the presence of 10 μm or 100 μm mg-Proto. mg-Proto was added in DmSO, and therefore the same amount of DmSO without mg-Proto was added for the control culture. total rna was extracted from the cells and Dna microarray analysis⁹ was performed to reveal the effect of Mg-Proto on the nuclear gene expression. Dashed lines showed two times changes in the level of gene expression. each value was obtained by the mean of two reproductions. Gray circles in the right plot indicate the genes annotated as chaperones. (B) northern analysis of the chaperone gene, HSP90. mg-Proto or hemin solved in DmSO were added to the C. merolae culture after the second 18 hours dark period of the 18 h-dark/6 h-light cycles with air-bubbled conditions, and the cultivation was continued for 1 hour in the dark. total rnas were prepared at the same timings as (a), and 10 μg of rnas were analyzed by northern hybridization. D18, after the second 18 hours dark period; D19, incubated for 1 hour with DmSO; other lanes are samples after 1 hour incubation with tetrapyrroles as indicated. Probe for the HSP90 gene was prepared by Pcr using following primers, HSP90F, tac atG aGc Gcc aaG aaG atc a; HSP90r, act tcc tcc atG Gca ctc tcG a. Labeling of probes and signal detection were performed by alkPhos direct labeling kit (Ge Healthcare uK, Buckinghamshire, england) and the image analyzer, LaS-3000 (Fuji Film, tokyo, Japan), respectively.