10
10
Supplementary Figure 5: Promoter analysis of
(A)
cmd
and (B)
trm-9
gene of
N. crassa
.
Gray boxes
showed the important regulatory sequences of gene and transcription start site (TSS) are indicated by using
arrows.
Supplementary Figure 6:
Expression studies of
cmd
gene in the presence of inhibitors. Fold change in
expression was calculated by 2
–ΔΔCt
method, using wild-type and β-tubulin as calibrator and endogenous
control respectively. Standard errors calculated from the data for two independent experiments are shown
using error bars.
Supplementary Figure 7:
Ergosterol is present in the
trm-9
mutant and
trm-9
nca-2
double mutant
strains. Profile of the sterols extracted from the wild-type,
nca-2
,
trm-9,
trm-9
nca-2,
and
erg-3
mutant strains were analysed by UV spectrophotometer.
Supplementary Figure 8:
(A) Calcium sensitivity, (B) development of aerial hyphae, and (C) UV survival.
(A) Ca
2+
sensitivity analysis of
wild-type,
trm-9,
nca-2
, and
trm-9
nca-2
double mutant strains.
Colony diameter (cm h
−1
) were measured at regular intervals and plotted against various concentrations of
CaCl
2
. Standard errors calculated from the data for three independent experiments are shown using error
bars. (B)
Aerial hyphae development of the wild-type,
trm-9
,
nca-2
, and
trm-9
nca-2
double mutant
strains on VSM agar media in test tube. The aerial hyphae growth of
trm-9
nca-2
double mutant strain
was less as compared to parental single mutants and wild-type strain. (C)
UV survival. Spot-test analysis
of wild-type,
trm-9,
nca-2,
and
trm-9
nca-2
double mutant and
upr-1
mutant strain grown on VG agar
at 30
C for 48 h in dark then illuminated for 24 h. UV survival assay was done essentially as described
previously (Deka et al., 2011).
Supplementary references:
Colot H.V., Park G., Turner G.E., Ringelberg C., Crew C.M., Litvinkova L., Weiss R.L., Borkovich K.A., Dunlap J.C., 2006, A high-throughput gene
knockout procedure for
Neurospora
reveals functions for multiple transcription factors, Proc Natl Acad Sci U S A 103:10352-10357
Deka R., Kumar R., Tamuli R., 2011,
Neurospora crassa
homologue of Neuronal Calcium Sensor-1 has a role in growth, calcium stress tolerance, and
ultraviolet survival, Genetica 139:885-894
