Table 3. Outcomes of modulating IMPα expression and transport function on exogenous dsRed2-PSPC1-positive nuclear foci.
| GFP | GFP-IMPα2-FL | GFP-IMPα2ΔIBB | GFP-IMPα2-ED | GFP-IMPα4-FL | GFP-IMPα4ΔIBB | GFP-IMPα6-FL | GFP-IMPα6ΔIBB | ||
|---|---|---|---|---|---|---|---|---|---|
| A) | Number of cells analysed - Total: 2530 | 180 | 228 | 241 | 446 | 317 | 386 | 513 | 219 |
| B) | DsRed2-PSPC1 nuclear foci - Total: 38657 | 2508 | 3392 | 2147 | 6218 | 10853 | 4339 | 5473 | 3745 |
| C) | Number of cells +VE for nuclear foci | 50 | 117 | 92 | 195 | 189 | 143 | 195 | 87 |
| D) | % Cells +VE for foci | 27.8% | 51.3% | 38.2% | 43.7% | 59.6% | 37% | 38% | 39.7% |
| 95% CI | (18.7 ↔ 36.8)% | (38.0 ↔ 64.6)% | (28.3 ↔ 48.1)% | (35.5 ↔ 51.9)% | (46.2 ↔ 73.0)% | (29.4 ↔ 44.7)% | (31.2 ↔ 44.8)% | (29.0 ↔ 50.5)% | |
| Odds Ratio (IMPα Normalised) | 1.000 | 0.586 | 0.737 | 1.000 | 0.399 | 1.000 | 1.075 | ||
| 95% CI | (Control) | (0.406 ↔ 0.846) | (0.535 ↔ 1.015) | (Control) | (0.294 ↔ 0.541) | (Control) | (0.777 ↔ 1.486) | ||
| Significance Value (Lg Reg) | p = 0.0042* | p = 0.0614 | p = 0.0000* | p = 0.6625 | |||||
| Mean per cell values (All cells) | GFP | GFP-IMPα2-FL | GFP-IMPα2ΔIBB | GFP-IMPα2-ED | GFP-IMPα4-FL | GFP-IMPα4ΔIBB | GFP-IMPα6-FL | GFP-IMPα6ΔIBB | |
| E) | GM Fn/c per cell - DsRed2-PSPC1 | 1.66 | 1.86 | 1.62 | 1.60 | 1.94 | 1.78 | 1.56 | 1.86 |
| 95% CI | (1.57 ↔ 1.76) | (1.75 ↔ 1.98) | (1.53 ↔ 1.72) | (1.54 ↔ 1.66) | (1.82 ↔ 2.06) | (1.70 ↔ 1.87) | (1.51 ↔ 1.62) | (1.72 ↔ 3.62) | |
| Odds Ratio (IMPα Normalised) | 1.000 | 0.873 | 0.860 | 1.000 | 0.922 | 1.000 | 1.191 | ||
| 95% CI | (Control) | (0.801 ↔ 0.951) | (0.798 ↔ 0.928) | (Control) | (0.859 ↔ 0.989) | (Control) | (1.105 ↔ 1.284) | ||
| Significance Value (Ln Reg) | p = 0.0019* | p = 0.0001* | p = 0.0235 | p = 0.0000* | |||||
| F) | GM intensity per cell - DsRed2-PSPC1 | 5.78 | 5.85 | 4.28 | 5.19 | 6.38 | 4.95 | 4.40 | 4.59 |
| 95% CI | (5.57 ↔ 6.00) | (5.66 ↔ 6.06) | (4.123 ↔ 4.434) | (4.99 ↔ 5.40) | (6.12 ↔ 6.65) | (4.76 ↔ 5.15) | (4.28 ↔ 4.53) | (4.38 ↔ 4.81) | |
| Odds Ratio (IMPα Normalised) | 1.000 | 0.730 | 0.887 | 1.000 | 0.777 | 1.000 | 1.042 | ||
| 95% CI | (Control) | (0.685 ↔ 0.779) | (0.838 ↔ 0.939) | (Control) | (0.737 ↔ 0.819) | (Control) | (0.986 ↔ 1.103) | ||
| Significance Value (Ln Reg) | p = 0.0000* | p = 0.0000* | p = 0.0000* | p = 0.144 | |||||
| G) | GM intensity per cell – GFP | 24.51 | 11.10 | 18.05 | 13.04 | 10.59 | 14.31 | 15.74 | 14.46 |
| 95% CI | (21.6 ↔ 27.9) | (10.30 ↔ 11.96) | (16.39 ↔ 19.89) | (12.29 ↔ 13.84) | (9.99 ↔ 11.22) | (13.25 ↔ 15.47) | (14.67 ↔ 16.88) | (13.05 ↔ 16.01) | |
| Odds Ratio (IMPα Normalised) | 1.000 | 1.627 | 1.175 | 1.000 | 1.352 | 1.000 | 0.919 | ||
| 95% CI | (Control) | (1.427 ↔ 1.855) | (1.047 ↔ 1.139) | (Control) | (1.214 ↔ 1.505) | (Control) | (0.819 ↔ 1.030) | ||
| Significance Value (Ln Reg) | p = 0.0000* | p = 0.0062* | p = 0.0000* | p = 0.1462 | |||||
| Mean per cell values (DsRed2-PSPC1 nuclear foci positive cells only) | GFP | GFP-IMPα2-FL | GFP-IMPα2ΔIBB | GFP-IMPα2-ED | GFP-IMPα4-FL | GFP-IMPα4ΔIBB | GFP-IMPα6-FL | GFP-IMPα6ΔIBB | |
| H) | GM number of foci (per cell) | 12.70 | 11.51 | 7.49 | 10.32 | 14.94 | 7.78 | 9.04 | 15.58 |
| 95% CI | (7.88 ↔ 20.48) | (9.04 ↔ 14.65) | (5.53 ↔ 10.13) | (8.39 ↔ 12.70) | (11.70 ↔ 19.07) | (5.96 ↔ 10.15) | (7.33 ↔ 11.14) | (11.30 ↔ 21.50) | |
| Odds Ratio (IMPα Normalised) | 1.000 | 0.651 | 0.897 | 1.000 | 0.521 | 1.000 | 1.725 | ||
| 95% CI | (Control) | (0.427 ↔ 0.992) | (0.629 ↔ 1.278) | (Control) | (0.372 ↔ 0.729) | (Control) | (1.167 ↔ 2.549) | ||
| Significance Value (Ln Reg) | p = 0.0462 | p = 0.5481 | p = 0.0001* | p = 0.0063* | |||||
| I) | GM ∑ foci volume (per cell) | 2.15 | 1.88 | 1.03 | 1.43 | 2.48 | 1.00 | 1.29 | 2.59 |
| 95% CI | (1.13 ↔ 4.08) | (1.37 ↔ 2.53) | (0.71 ↔ 1.51) | (1.10 ↔ 1.86) | (1.79 ↔ 3.42) | (0.71 ↔ 1.42) | (0.97 ↔ 1.71) | (1.71 ↔ 3.91) | |
| Odds Ratio (IMPα Normalised) | 1.000 | 0.550 | 0.761 | 1.000 | 0.405 | 1.000 | 2.003 | ||
| 95% CI | (Control) | (0.316 ↔ 0.956) | (0.478 ↔ 1.211) | (Control) | (0.261 ↔ 0.628) | (Control) | (1.200 ↔ 3.343) | ||
| Significance Value (Ln Reg) | p = 0.0342 | p = 0.2490 | p = 0.0001* | p = 0.0080 | |||||
| J) | GM ∑ foci DsRed2-PSPC1 intensity (per cell) | 8724 | 7704 | 3921 | 5499 | 10240 | 3838 | 5102 | 10400 |
| 95% CI | (4397 ↔ 17310) | (5476 ↔ 10840) | (2602 ↔ 5908) | (4152 ↔ 7283) | (7250 ↔ 14460) | (2649 ↔ 5562) | (3744 ↔ 6954) | (6684 ↔ 16190) | |
| Odds Ratio (IMPα Normalised) | 1.000 | 0.509 | 0.714 | 1.000 | 0.375 | 1.000 | 2.039 | ||
| 95% CI | (Control) | (0.281 ↔ 9.214) | (0.434 ↔ 1.175) | (Control) | (0.234 ↔ 6.012) | (Control) | (1.177 ↔ 3.532) | ||
| Significance Value (Ln Reg) | p = 0.0259 | p = 0.1850 | p = 0.0001* | p = 0.0112 | |||||
| Mean values per DsRed2-PSPC1 nuclear foci | GFP | GFP-IMPα2-FL | GFP-IMPα2ΔIBB | GFP-IMPα2-ED | GFP-IMPα4-FL | GFP-IMPα4ΔIBB | GFP-IMPα6-FL | GFP-IMPα6ΔIBB | |
| K) | GM foci volume (per foci) | 0.215 | 0.162 | 0.159 | 0.155 | 0.229 | 0.179 | 0.197 | 0.180 |
| 95% CI | (0.207 ↔ 0.224) | (0.157 ↔ 0.167) | (0.152 ↔ 0.166) | (0.151 ↔ 0.159) | (0.225 ↔ 0.234) | (0.174 ↔ 0.185) | (0.192 ↔ 0.202) | (0.174 ↔ 0.185) | |
| Odds Ratio (IMPα Normalised) | 1.000 | 0.980 | 0.956 | 1.000 | 0.783 | 1.000 | 0.911 | ||
| 95% CI | (Control) | (0.710 ↔ 1.353) | (0.779 ↔ 1.175) | (Control) | (0.629 ↔ 0.975) | (Control) | (0.709 ↔ 1.172) | ||
| Significance Value (GEE) | p = 0.9026 | p = 0.6707 | p = 0.0290 | p = 0.469 | |||||
| L) | GM foci DsRed2-PSPC1 voxel intensity (per foci) | 99.89 | 95.18 | 97.14 | 92.37 | 102.8 | 95.52 | 102.1 | 97.08 |
| 95% CI | (98.84 ↔ 100.9) | (94.42 ↔ 95.94) | (95.95 ↔ 98.34) | (91.83 ↔ 92.92) | (102.3 ↔ 103.4) | (94.76 ↔ 96.29) | (101.3 ↔ 102.9) | (96.22 ↔ 97.94) | |
| Odds Ratio (IMPα Normalised) | 1.000 | 1.023 | 0.971 | 1.000 | 0.929 | 1.000 | 0.951 | ||
| 95% CI | (Control) | (0.931 ↔ 1.12) | (0.924 ↔ 1.02) | (Control) | (0.861 ↔ 1.002) | (Control) | (0.880 ↔ 1.026) | ||
| Significance Value (GEE) | p = 0.637 | p = 0.232 | p = 0.0565 | p = 0.1947 | |||||
| M) | GM ∑ foci DsRed2-PSPC1 intensity (per foci) | 885 | 614 | 613 | 577 | 970 | 694 | 820 | 707 |
| 95% CI | (845 ↔ 928) | (590 ↔ 638) | (581 ↔ 647) | (560 ↔ 594) | (946 ↔ 994) | (669 ↔ 720) | (793 ↔ 848) | (679 ↔ 736) | |
| Odds Ratio (IMPα Normalised) | 1.000 | 1.00 | 0.94 | 1.000 | 0.716 | 1.000 | 0.862 | ||
| 95% CI | (Control) | (0.649 ↔ 1.55) | (0.726 ↔ 1.22) | (Control) | (0.532 ↔ 0.963) | (Control) | (0.617 ↔ 1.205) | ||
| Significance Value (GEE) | p = 0.991 | p = 0.6383 | p = 0.0270 | p = 0.385 | |||||
The analysed cell numbers for each GFP-tagged IMPα group, all co-transfected with DsRed2-PSPC1, the number of detected DsRed2-PSPC1-positive nuclear foci and proportion of cells determined to contain DsRed2-PSPC1 nuclear foci (detected by DsRed2-PSPC1 fluorescence) are presented. Samples were assessed on a per cell or per DsRed2-PSPC1 nuclear foci basis, with geometric means (GM) and 95% confidence intervals (95% CI) calculated. To determine significant differences between groups, a logistic regression (Lg Reg) model was used for DsRed2-PSPC1 foci positive/negative cells, linear regression (Ln Reg) models were used for per cell data and generalised estimating equations (GEE) were used for per DsRed2-PSPC1 nuclear foci data. Comparative significance values using IMPα-FL as the reference groups (set at 1.000) are shown. Using Bonferroni correction, the significance threshold was reassigned from ≤0.05 to ≤0.0063 (0.05 ÷ 8 experimental groups), with those outcomes below the threshold indicated (*). Further details are provided in Fig. 2B with additional samples and analysis parameters included in Supplementary Tables S4 and S7.