TABLE 6.
CYP51 and efflux transporter gene mRNA expression levels in the mutant strains assessed by real-time RT-PCR
| Strain | cDNA levela
|
cDNA level
|
||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
|
Afumdr1
|
Afummdr2
|
Afumdr3
|
Afumdr4
|
AtrF
|
CYP51A
|
CYP51B
|
||||||||
| Const. expres.b | Itra treatedb | Const. expres. | Itra treated | Const. expres. | Itra treated | Const. expres. | Itra treated | Const. expres. | Itra treated | Const. expres. | Itra treated | Const. expres. | Itra treated | |
| CEA17 | 1.00 (1.00-1.00) | 3.67 (317-4.17) | 1.00 (1.00-1.00) | 0.33 (0.30-0.37) | 1.00 (0.75-1.25) | 1.50 (1.25-1.75) | 1.00 (0.97-1.03) | 23.85 (23.38-24.31)b | 1.00 (0.80-1.20) | 0.20 (0.20-0.20)b | 1.00 (1.00-1.00) | 7.20 (6.72-7.69)b | 1.00 (0.97-1.03) | 4.27 (3.94-4.61)b |
| G10-1-50A | 0.50 (0.50-0.50) | 12.33 (11.67-13.00) | 1.00 (0.98-1.02) | 2.89 (2.94-2.84) | 13.25 (13.00-13.50) | 20.17 (18.56-21.72) | 5.03 (4.85-5.21) | 11.55 (11.53-11.57) | 1.00 (1.00-1.00) | 17.60 (17.40-17.80) | 1.69 (1.67-1.70) | 2.82 (2.69-2.96) | 1.18 (1.08-1.28) | 2.31 (2.34-2.28) |
| G10-1-50B | 5.00 (5.00-5.00) | 2.53 (2.50-2.57) | 1.78 (1.64-1.91) | 3.38 (3.33-3.41) | 174.50 (160.75-188.25) | 3.56 (3.55-3.56) | 28.46 (27.18-29.74) | 6.25 (6.09-6.40) | 7.60 (6.80-8.40) | 5.61 (6.24-5.10) | 0.72 (0.70-0.74) | 7.13 (7.21-7.05) | 0.55 (0.52-0.57) | 4.17 (3.89-4.43) |
| G10-2-50A | 0.83 (0.83-0.83) | 6.40 (3.80-9.00) | 0.89 (0.88-0.90) | 5.38 (3.03-7.67) | 6.50 (6.25-6.75) | 60.58 (33.40-85.74) | 2.82 (2.79-2.85) | 35.69 (21.06-50.05) | 0.60 (0.60-0.60) | 35.00 (22.00-48.00) | 1.19 (1.07-1.30) | 3.70 (2.52-4.69) | 0.55 (0.54-0.55) | 2.83 (1.68-3.95) |
| G10-3-50A | 0.67 (0.67-0.67) | 5.25 (5.25-5.25) | 0.89 (0.87-0.91) | 5.50 (4.33-6.61) | 40.75 (37.25-44.25) | 0.15 (0.13-0.16) | 7.87 (7.69-8.05) | 2.49 (2.53-2.46) | 2.40 (2.40-2.40) | 0.25 (0.25-0.25) | 1.65 (1.65-1.65) | 4.06 (3.85-4.26) | 1.18 (1.17-1.19) | 2.54 (2.15-2.92) |
| H05-3-10 | 0.18 (0.17-0.18) | 0.29 (0.30-0.28) | 0.16 (0.16-0.16) | 0.41 (0.13-0.68) | 0.02 (0.02-0.02) | 0.60 (0.04-1.13) | 0.52 (0.10-0.94) | 0.05 (0.19-0.04) | 2.22 (2.19-2.26) | 0.18 (0.18-0.19) | 4.48 (4.13-4.83) | 2.43 (2.52-2.35) | 1.08 (0.94-1.22) | 0.60 (0.58-0.62) |
| H10-1-50A | 0.83 (0.83-0.83) | 6.40 (6.00-6.80) | 1.78 (1.07-2.49) | 1.88 (3.01-1.39) | 324.25 (315.25-333.25) | 1.09 (1.03-1.14) | 7.97 (7.90-8.05) | 8.77 (7.77-9.75) | 2.00 (1.80-2.20) | 12.60 (13.44-11.91) | 1.17 (1.09-1.24) | 3.63 (3.88-3.42) | 0.82 (0.81-0.83) | 2.22 (1.38-3.04) |
| H10-2-50A | 0.83 (0.83-0.83) | 4.00 (3.80-4.20) | 0.67 (0.62-0.71) | 3.33 (3.41-3.27) | 2.00 (2.00-2.25) | 214.38 (201.13-202.33) | 2,769 (2,717-2,821) | 15.52 (14.71-16.30) | 0.20 (0.20-0.20) | 108.0 (104.0-112.0) | 1.65 (1.63-1.67) | 2.75 (2.77-2.73) | 0.82 (0.82-0.82) | 3.00 (2.89-3.11) |
| H10-3-50A | 5.00 (4.83-5.17) | 0.53 (0.52-0.55) | 28.00 (27.83-28.17) | 0.03 (0.03-0.03) | 178.25 (118.25-238.25) | 0.29 (0.41-0.23) | 170.87 (168.97-172.77) | 0.06 (0.06-0.06) | 17.20 (15.40-19.00) | 0.21 (0.22-0.20) | 0.26 (0.24-0.28) | 24.71 (25.62-23.93) | 1.45 (1.41-1.50) | 1.50 (1.54-1.47) |
| H10-1-50B | 0.18 (0.18-0.18) | 0.71 (0.68-0.74) | 0.10 (0.09-0.11) | 1.49 (1.60-1.40) | 0.0017 (0.0011-0.0023) | 74.75 (107.10-58.98) | 0.18 (0.16-0.19) | 4.13 (3.89-4.32) | 3.38 (3.19-3.58) | 2.57 (2.53-2.60) | 6.05 (5.84-6.25) | 1.10 (1.09-1.12) | 0.54 (0.53-0.54) | 0.67 (0.53-0.80) |
| RIT13 | 0.33 (0.33-0.33) | 16.00 (14.50-17.50) | 0.89 (0.89-0.89) | 38.25 (37.70-38.80) | 23.75 (20.75-26.75) | 258.33 (257.73-258.79) | 7.41 (7.31-7.51) | 249.3 (237.0-261.30) | 1.80 (1.60-2.00) | 162.22 (180.00-148.00) | 1.20 (1.09-1.33) | 2.81 (2.50-3.13) | 0.64 (0.64-0.64) | 2.86 (2.70-3.01) |
a
Const. expres., constitutive expression, cDNA levels were calculated relative to that of the untreated parental strain, CEA17. The data represent the averages of three replicates, with the ranges shown in parentheses.
b
Values were obtained by using the cDNA copy number of each untreated strain as the baseline for strains in the presence of drug following normalization based on the GPDH cDNA copy number. Cells were grown in the presence of 10 ng of itraconazole (Itra)/ml (CEA17) or 5 μg of itraconazole/ml for 24 h. Ranges are shown in parentheses.