Table 2. Outcomes of modulating IMPα expression and transport function on endogenous SFPQ-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: 2258 | 566 | 192 | 301 | 143 | 173 | 299 | 273 | 311 |
| B) | SFPQ nuclear foci - Total: 14750 | 4570 | 1529 | 1666 | 845 | 1787 | 1297 | 2123 | 933 |
| C) | Number of cells + VE for nuclear foci | 460 | 161 | 174 | 84 | 158 | 180 | 234 | 128 |
| D) | % Cells +VE for foci | 81.3% | 83.9% | 57.8% | 58.7% | 91.3% | 60.2% | 85.7% | 41.2% |
| 95% CI | (64.1 ↔ 98.4)% | (51.6 ↔ 116.1)% | (44.6 ↔ 71.0)% | (39.2 ↔ 78.3)% | (43.0 ↔ 139.7)% | (46.3 ↔ 74.1)% | (56.7 ↔ 114.8)% | (31.9 ↔ 50.5)% | |
| Odds Ratio (IMPα Normalised) | 1.000 | 0.264 | 0.274 | 1.000 | 0.144 | 1.000 | 0.117 | ||
| 95% CI | (Control) | (0.169 ↔ 0.413) | (0.165 ↔ 0.456) | (Control) | (0.081 ↔ 0.256) | (Control) | (0.078 ↔ 0.175) | ||
| Significance Value (Lg Reg) | p = 0.0000* | p = 0.0000* | p = 0.0000* | p = 0.0000* | |||||
| 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 - SFPQ-A546 | 6.24 | 4.75 | 3.21 | 2.80 | 5.49 | 3.93 | 5.32 | 2.66 |
| 95% CI | (6.01 ↔ 6.47) | (4.43 ↔ 5.09) | (2.93 ↔ 3.50) | (2.57 ↔ 3.06) | (5.21 ↔ 5.79) | (3.70 ↔ 4.19) | (5.05 ↔ 5.62) | (2.44 ↔ 2.90) | |
| Odds Ratio (IMPα Normalised) | 1.000 | 0.675 | 0.590 | 1.000 | 0.716 | 1.000 | 0.500 | ||
| 95% CI | (Control) | (0.609 ↔ 0.749) | (0.521 ↔ 0.668) | (Control) | (0.643 ↔ 0.797) | (Control) | (0.456 ↔ 0.549) | ||
| Significance Value (Ln Reg) | p = 0.0000* | p = 0.0000* | p = 0.0000* | p = 0.0000* | |||||
| F) | GM intensity per cell - SFPQ-A546 | 7.13 | 9.22 | 6.70 | 11.04 | 9.68 | 7.59 | 8.55 | 5.34 |
| 95% CI | (6.95 ↔ 7.33) | (8.70 ↔ 9.77) | (6.29 ↔ 7.14) | (9.90 ↔ 12.32) | (9.22 ↔ 10.16) | (7.24 ↔ 9.95) | (8.21 ↔ 8.90) | (5.07 ↔ 5.63) | |
| Odds Ratio (IMPα Normalised) | 1.000 | 0.727 | 1.198 | 1.000 | 0.784 | 1.000 | 0.625 | ||
| 95% CI | (Control) | (0.672 ↔ 0.786) | (1.091 ↔ 1.315) | (Control) | (0.723 ↔ 0.849) | (Control) | (0.583 ↔ 0.670) | ||
| Significance Value (Ln Reg) | p = 0.0000* | p = 0.0002* | p = 0.0000* | p = 0.0000* | |||||
| G) | GM intensity per cell – GFP | 13.38 | 9.34 | 11.91 | 8.40 | 9.42 | 12.50 | 12.91 | 11.00 |
| 95% CI | (12.76 ↔ 14.04) | (8.63 ↔ 10.11) | (11.20 ↔ 12.67) | (7.76 ↔ 9.10) | (8.67 ↔ 10.24) | (11.62 ↔ 13.38) | (11.66 ↔ 14.28) | (10.17 ↔ 11.90) | |
| Odds Ratio (IMPα Normalised) | 1.000 | 1.276 | 0.900 | 1.000 | 1.326 | 1.000 | 0.852 | ||
| 95% CI | (Control) | (1.138 ↔ 1.429) | (0.785 ↔ 1.031) | (Control) | (1.179 ↔ 1.492) | (Control) | (0.769 ↔ 0.944) | ||
| Significance Value (Ln Reg) | p = 0.0000* | p = 0.1286 | p = 0.0000* | p = 0.0022* | |||||
| Mean per cell values (SFPQ-A546 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) | 7.61 | 6.25 | 7.10 | 7.50 | 8.13 | 5.14 | 6.85 | 5.25 |
| 95% CI | (7.07 ↔ 8.20) | (5.39 ↔ 7.26) | (6.24 ↔ 8.07) | (6.23 ↔ 9.01) | (7.08 ↔ 9.34) | (4.54 ↔ 5.82) | (6.17 ↔ 7.60) | (4.51 ↔ 6.11) | |
| Odds Ratio (IMPα Normalised) | 1.000 | 1.135 | 1.199 | 1.000 | 0.632 | 1.000 | 0.766 | ||
| 95% CI | (Control) | (0.945 ↔ 1.363) | (0.957 ↔ 1.501) | (Control) | (0.527 ↔ 0.759) | (Control) | (0.637 ↔ 0.921) | ||
| Significance Value (Ln Reg) | p = 0.1751 | p = 0.1152 | p = 0.0000* | p = 0.0046* | |||||
| I) | GM ∑ foci volume (per cell) | 2.68 | 1.96 | 2.44 | 2.33 | 3.11 | 1.16 | 1.99 | 1.38 |
| 95% CI | (2.33 ↔ 3.07) | (1.51 ↔ 2.54) | (1.94 ↔ 3.06) | (1.70 ↔ 3.20) | (2.48 ↔ 3.89) | (0.95 ↔ 1.41) | (1.65 ↔ 2.41) | (1.05 ↔ 1.80) | |
| Odds Ratio (IMPα Normalised) | 1.000 | 1.247 | 1.1931 | 1.000 | 0.3731 | 1.000 | 0.691 | ||
| 95% CI | (Control) | (0.904 ↔ 1.722) | (0.802 ↔ 1.774) | (Control) | (0.271 ↔ 0.515) | (Control) | (0.499 ↔ 0.955) | ||
| Significance Value (Ln Reg) | p = 0.1792 | p = 0.3835 | p = 0.0000* | p = 0.0254 | |||||
| J) | GM ∑ foci SFPQ-A546 intensity (per cell) | 9422 | 7276 | 9306 | 9159 | 12102 | 4343 | 7557 | 5052 |
| 95% CI | (8159 ↔ 10879) | (5567 ↔ 9509) | (7313 ↔ 11843) | (6604 ↔ 12703) | (9600 ↔ 15255) | (3543 ↔ 5322) | (6200 ↔ 9212) | (3803 ↔ 6712) | |
| Odds Ratio (IMPα Normalised) | 1.000 | 1.279 | 1.259 | 1.000 | 0.359 | 1.000 | 0.669 | ||
| 95% CI | (Control) | (0.914 ↔ 1.790) | (0.833 ↔ 1.903) | (Control) | (0.257 ↔ 0.502) | (Control) | (0.477 ↔ 0.973) | ||
| Significance Value (Ln Reg) | p = 0.1511 | p = 0.2753 | p = 0.0000* | p = 0.0195 | |||||
| Mean values per SFPQ-A546 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) | 0.318 | 0.329 | 0.317 | 0.286 | 0.320 | 0.202 | 0.264 | 0.264 |
| 95% CI | (0.307 ↔ 0.329) | (0.309 ↔ 0.349) | (0.300 ↔ 0.308) | (0.266 ↔ 0.308) | (0.303 ↔ 0.337) | (0.191 ↔ 0.214) | (0.252 ↔ 0.277) | (0.246 ↔ 0.283) | |
| Odds Ratio (IMPα Normalised) | 1.000 | 0.965 | 0.871 | 1.000 | 0.632 | 1.000 | 0.998 | ||
| 95% CI | (Control) | (0.773 ↔ 1.204) | (0.693 ↔ 1.094) | (Control) | (0.538 ↔ 0.741) | (Control) | (0.842 ↔ 1.184) | ||
| Significance Value (GEE) | p = 0.750 | p = 0.234 | p = 0.0000* | p = 0.9837 | |||||
| L) | GM foci SFPQ-A546 voxel intensity (per foci) | 89.56 | 87.21 | 89.01 | 89.26 | 90.16 | 87.65 | 87.48 | 88.31 |
| 95% CI | (89.17 ↔ 89.95) | (86.57 ↔ 87.85) | (88.32 ↔ 89.71) | (88.42 ↔ 90.11) | (89.51 ↔ 90.82) | (87.08 ↔ 88.22) | (86.97 ↔ 87.99) | (87.60 ↔ 89.03) | |
| Odds Ratio (IMPα Normalised) | 1.000 | 1.02 | 1.02 | 1.000 | 0.972 | 1.000 | 1.009 | ||
| 95% CI | (Control) | (0.998 ↔ 1.05) | (0.999 ↔ 1.05) | (Control) | (0.953 ↔ 0.991) | (Control) | (0.991 ↔ 1.03) | ||
| Significance Value (GEE) | p = 0.0736 | p = 0.0582 | p = 0.0045* | p = 0.3070 | |||||
| M) | GM ∑ foci SFPQ-A546 intensity (per foci) | 1107 | 1202 | 1188 | 1071 | 1204 | 728 | 964 | 963 |
| 95% CI | (1068 ↔ 1149) | (1126 ↔ 1282) | (1117 ↔ 1263) | (988 ↔ 1160) | (1135 ↔ 1278) | (684 ↔ 775) | (914 ↔ 1017) | (891 ↔ 1041) | |
| Odds Ratio (IMPα Normalised) | 1.000 | 0.993 | 0.891 | 1.000 | 0.604 | 1.000 | 0.999 | ||
| 95% CI | (Control) | (0.782 ↔ 1.260) | (0.701 ↔ 1.132) | (Control) | (0.507 ↔ 0.720) | (Control) | (0.835 ↔ 1.196) | ||
| Significance Value (GEE) | p = 0.952 | p = 0.344 | p = 0.0000* | p = 0.9931 | |||||
The analysed cell numbers for each GFP-tagged IMPα transfection group, the number of detected SFPQ-positive nuclear foci and proportion of cells determined to contain SFPQ nuclear foci (detected by indirect SFPQ immunofluorescence with an Alexa Fluor 546 [A546] secondary antibody) are presented. Samples were assessed on a per cell or per SFPQ 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 SFPQ foci positive/negative cells, linear regression (Ln Reg) models were used for per cell data and generalised estimating equations (GEE) were used for per 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. 2C with additional samples and analysis parameters included in Supplementary Tables S3 and S6.