Table 1.
CD82 expression effects in neonatal and adult O4+ cells after 7 d in vitro in the presence or absence of HGF
|
Control–EGFP |
CD82–IRES–EGFP |
shRNA–ZsGreen |
||||
|---|---|---|---|---|---|---|
| −HGF | +HGF | −HGF | +HGF | −HGF | +HGF | |
| Neonatal | ||||||
| Process number | ||||||
| 0 | 23 (26.7%) | 37 (61.7%) | 0 (0%) | 0 (0%) | 42 (84%) | 58 (78.4%) |
| <4 | 47 (54.7%) | 23 (38.3%) | 6 (11.1%) | 5 (11.4%) | 8 (16%) | 16 (21.6%) |
| >5 | 16 (18.6%) | 0 (0%) | 48 (88.9%) | 39 (88.6%) | 0 (0%) | 0 (0%) |
| Marker expression | ||||||
| NG2 | 3 of 56 (5.4%) | 11 of 47 (23.4%) | 0 (0%) | 0 (0%) | 25 of 40 (62.5%) | 20 of 58 (34.5%) |
| PDGFRα | 7 of 49 (14.3%) | 13 of 42 (30.9%) | 0 (0%) | 0 (0%) | 41 of 50 (82%) | 42 of 51 (82.4%) |
| O1 | 31 of 43 (72.1%) | 26 of 55 (47.3%) | 29 of 30 (96.7%) | 35 of 35 (100%) | 0 (0%) | 0 (0%) |
| MBP | 5 of 47 (10.6%) | 0 (0%) | 40 of 56 (71.4%) | 32 of 60 (53.3%) | 0 (0%) | 0 (0%) |
| Adult | ||||||
| Process number | ||||||
| 0 | 16 (11.8%) | 14 (18.4%) | 0 (0%) | 0 (0%) | 5 (12.2%) | 23 (56.1%) |
| <4 | 44 (32.3%) | 23 (30.3%) | 14 (29.2%) | 12 (28.6%) | 28 (68.3%) | 18 (43.9%) |
| >5 | 76 (55.9%) | 39 (51.3%) | 34 (70.8%) | 30 (71.4%) | 8 (19.5%) | 0 (0%) |
| Marker expression | ||||||
| NG2 | 0 (0%) | 0 (0%) | 0 (0%) | 0 (0%) | 0 (0%) | 14 of 40 (35%) |
| PDGFRα | 0 (0%) | 0 (0%) | 0 (0%) | 0 (0%) | 44 of 54 (81.5%) | 26 of 44 (59.1%) |
| O1 | 52 of 52 (100%) | 49 of 49 (100%) | 44 of 44 (100%) | 37 of 37 (100%) | 24 of 64 (37.5%) | 0 (0%) |
| MBP | 20 of 42 (47.6%) | 14 of 55 (25.5%) | 25 of 32 (78.1%) | 28 of 58 (48.3%) | 0 (0%) | 0 (0%) |
Five representative 40× fields from two separate experiments were photographed per marker. Green and double-positive cells and their number of processes were manually counted. Statistical analysis was performed using χ2 test. There are statistically significant differences in process numbers between all populations in the presence and absence of HGF, with the exception of control–EGFP+ adult cells. There are statistically significant differences in marker expression between all populations in the presence and absence of HGF, with the exception of NG2, PDGFRα (0% expected and observed cells), and O1 expression between adult CD82–IRES–EGFP+ and control–EGFP+ cells. Number of processes: neonatal cells, x2CD82–IRES–EGFP+/control–EGFP+ (−HGF) = 9.098e−72, x2shRNA–ZsGreen+/control–EGFP+ (−HGF) = 2.056e−37, x2CD82–IRES–EGFP+/control–EGFP+ (+HGF) = 2.773e−19, x2shRNA–ZsGreen+/control–EGFP+(+HGF) = 0.000592, x2control–EGFP+ (±HGF) = 8.523e−16; adult cells, x2CD82–IRES–EGFP+/control–EGFP+ (−HGF) = 0.000324, x2shRNA–ZsGreen+/control–EGFP+ (−HGF) = 1.372e−14, x2CD82–IRES–EGFP+/control–EGFP+ (+HGF) = 1.878e−06, x2shRNA–ZsGreen+/control–EGFP+ (+HGF) = 5.774e−30, x2control–EGFP+ (±HGF) = 0.12283 (x20.05 = 0.103, df = 2). Marker expression: neonatal cells, CD82–IRES–EGFP+/control–EGFP+, x2NG2+ = 8.025e−08, x2PDGFRα+ = 1.779e−11, x2O1+ = 2.568e−16, x2MBP+ = 1.177e−24; shRNA–ZsGreen+/control–EGFP+, x2NG2+ = 1.805e−134, x2PDGFRα+ = 2.292e−90, x2O1+ = 8.56e−28, x2MBP+ = 0.00113, x2control–EGFP+ (±HGF) = 3.431e−21; adult cells, CD82–IRES–EGFP+/control–EGFP+, x2O1+ = 1, x2MBP+ = 2.63371e−10; shRNA–ZsGreen+/control–EGFP+, x2NG2+ = 0, x2PDGFRα+ = 1.96793e−32, x2O1+ = 4.26804e−32, x2MBP+ = 1.23245e−17, x2control–EGFP+ (±HGF) = 0.00136 (x20.05 = 0.00393, df = 1).