Table 3.
Statistical comparison of normalization schemes
Experiment | No normalization | Linear normalization | Global lowess | P-lowess | S-lowess | LIN | OLIN | OSLIN | |
Intensity-dependent cor(M, ![]() |
SW480/620 | 0.50 | 0.50 | 0.01 | -0.01 | -0.01 | 0.04 | 0.00 | 0.00 |
Intensity-dependent cor(M, ![]() |
Apo AI | 0.47 | 0.47 | 0.06 | 0.05 | 0.05 | 0.09 | 0.00 | 0.00 |
Spatial cor(M, ![]() |
SW480/620 | 0.53 | 0.53 | 0.56 | 0.34 | 0.32 | 0.27 | 0.07 | 0.08 |
Spatial cor(M, ![]() |
Apo AI | 0.58 | 0.58 | 0.59 | 0.41 | 0.38 | 0.43 | 0.15 | 0.15 |
Mean pairwise cor(M) | SW480/620 | 0.46 | 0.46 | 0.50 | 0.59 | 0.59 | 0.63 | 0.67 | 0.64 |
Var(M) | SW480/620 | 927.1 | 658.7 | 455.4 | 216.3 | 213.0 | 205.7 | 163.0 | 186.5 |
Number of significant genes | SW480/620 | 26 | 71 | 51 | 75 | 88 | 94 | 99 | 129 |
Average cor(MqPCR, Mma) | Fibroblast | 0.82 | 0.81 | 0.82 | 0.82 | 0.81 | 0.81 | 0.84 | 0.88 |
Intensity-dependent correlation (intensity-dependent cor(M, )) describes the correlation between the log ratio M of the spot and the median value of M within a symmetrical neighborhood of 50 spots on the intensity scale. Spatial correlation (spatial cor(M,
)) describes the correlation between the log ratio M of the spot and the median value of M within a neighborhood defined by a window of 5 × 5 spots. To ensure independence, M of the spot was not included in the median
of the neighborhood. For the calculation of mean pairwise correlation of slides, spots with intensity A < 11.6 were excluded. The significance of differential gene expression was examined by a one-sample t-test with the null hypothesis of mean log ratio M = 0. Duplicated spots on SW480/620 arrays were treated as independent measurements producing a maximum of eight observations per gene. Genes were detected as significantly differentially expressed if their Bonferroni adjusted p-values were smaller than 0.01. For the fibroblast experiment, the average Pearson correlation between qPCR-based logged fold changes MqPCR and microarray-based logged fold changes Mma of the genes for IL-8, COX2, Mast, B4-2 and actin is shown.