Table 7.
Analysis of reference gene expression variability in the femoral muscle during the development of obesity.
| NormFinder | GeNorm | BestKeeper | Delta-Ct | Consensus | ||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Genes | Stability value | Genes | Stability value | Genes | SD | Genes | SD | Genes | Geometric mean of ranking values | |
| 4 w NFD/HFD (n = 8/8) |
PPIA | 0.024 | HPRT | 0.196 | RPLP0 | 0.253 | RPLP0 | 0.328 | RPLP0 | 2.11 |
| HPRT | 0.035 | TBP | 0.196 | PPIA | 0.304 | HPRT | 0.374 | HPRT | 2.21 | |
| TBP | 0.045 | YWHAZ | 0.208 | TBP | 0.312 | PPIA | 0.379 | PPIA | 2.34 | |
| YWHAZ | 0.056 | RPLP0 | 0.216 | 18S | 0.321 | TBP | 0.385 | TBP | 2.91 | |
| RPLP0 | 0.066 | PPIA | 0.238 | GAPDH | 0.333 | YWHAZ | 0.386 | YWHAZ | 4.53 | |
| B2M | 0.087 | ACTB | 0.262 | HPRT | 0.343 | ACTB | 0.422 | ACTB | 6.70 | |
| ACTB | 0.106 | GAPDH | 0.291 | YWHAZ | 0.344 | B2M | 0.432 | 18S | 6.90 | |
| 18S | 0.114 | B2M | 0.315 | ACTB | 0.367 | GAPDH | 0.483 | GAPDH | 7.09 | |
| GAPDH | 0.132 | 18S | 0.334 | B2M | 0.379 | 18S | 0.496 | B2M | 7.42 | |
| UBC | 0.406 | UBC | 0.504 | UBC | 1.056 | UBC | 1.082 | UBC | 10.00 | |
| 8 w NFD/HFD (n = 8/8) |
HPRT | 0.127 | HPRT | 0.191 | 18S | 0.499 | PPIA | 0.355 | HPRT | 2.43 |
| TBP | 0.135 | GAPDH | 0.191 | B2M | 0.565 | RPLP0 | 0.368 | PPIA | 3.22 | |
| GAPDH | 0.148 | PPIA | 0.230 | YWHAZ | 0.619 | YWHAZ | 0.394 | YWHAZ | 3.46 | |
| YWHAZ | 0.159 | YWHAZ | 0.260 | RPLP0 | 0.633 | B2M | 0.396 | GAPDH | 3.98 | |
| ACTB | 0.166 | TBP | 0.280 | HPRT | 0.685 | ACTB | 0.405 | RPLP0 | 4.28 | |
| PPIA | 0.168 | RPLP0 | 0.295 | PPIA | 0.693 | GAPDH | 0.406 | B2M | 4.60 | |
| RPLP0 | 0.213 | B2M | 0.308 | GAPDH | 0.715 | HPRT | 0.407 | TBP | 5.03 | |
| B2M | 0.334 | ACTB | 0.320 | TBP | 0.728 | TBP | 0.444 | 18S | 5.20 | |
| 18S | 0.464 | 18S | 0.352 | ACTB | 0.824 | 18S | 0.575 | ACTB | 6.51 | |
| UBC | 1.027 | UBC | 0.525 | UBC | 1.449 | UBC | 1.034 | UBC | 10.00 | |
| 12 w NFD/HFD (n = 8/8) |
TBP | 0.085 | YWHAZ | 0.237 | TBP | 0.370 | PPIA | 0.332 | YWHAZ | 2.63 |
| HPRT | 0.119 | 18S | 0.237 | YWHAZ | 0.376 | RPLP0 | 0.402 | TBP | 2.71 | |
| PPIA | 0.127 | PPIA | 0.255 | HPRT | 0.383 | ACTB | 0.406 | PPIA | 2.71 | |
| B2M | 0.134 | GAPDH | 0.268 | B2M | 0.385 | YWHAZ | 0.408 | HPRT | 4.41 | |
| GAPDH | 0.165 | RPLP0 | 0.280 | 18S | 0.404 | B2M | 0.415 | 18S | 4.86 | |
| YWHAZ | 0.182 | ACTB | 0.295 | PPIA | 0.423 | TBP | 0.430 | B2M | 5.03 | |
| 18S | 0.247 | HPRT | 0.320 | ACTB | 0.428 | GAPDH | 0.443 | RPLP0 | 5.18 | |
| ACTB | 0.292 | B2M | 0.335 | RPLP0 | 0.460 | 18S | 0.450 | ACTB | 5.63 | |
| RPLP0 | 0.333 | TBP | 0.386 | GAPDH | 0.472 | HPRT | 0.457 | GAPDH | 5.96 | |
| UBC | 0.875 | UBC | 0.608 | UBC | 1.037 | UBC | 1.183 | UBC | 10.00 | |
| 16 w NFD/HFD (n = 8/8) |
RPLP0 | 0.136 | RPLP0 | 0.217 | PPIA | 0.273 | RPLP0 | 0.339 | RPLP0 | 1.68 |
| B2M | 0.176 | YWHAZ | 0.217 | TBP | 0.515 | YWHAZ | 0.414 | YWHAZ | 2.63 | |
| YWHAZ | 0.179 | B2M | 0.255 | 18S | 0.560 | B2M | 0.414 | B2M | 3.08 | |
| GAPDH | 0.188 | ACTB | 0.273 | YWHAZ | 0.592 | GAPDH | 0.450 | PPIA | 4.56 | |
| ACTB | 0.195 | GAPDH | 0.294 | B2M | 0.596 | ACTB | 0.469 | GAPDH | 4.68 | |
| PPIA | 0.269 | 18S | 0.331 | GAPDH | 0.614 | HPRT | 0.513 | 18S | 5.45 | |
| 18S | 0.311 | HPRT | 0.366 | UBC | 0.626 | 18S | 0.521 | ACTB | 5.62 | |
| TBP | 0.318 | TBP | 0.396 | RPLP0 | 0.634 | PPIA | 0.538 | TBP | 5.83 | |
| HPRT | 0.349 | PPIA | 0.440 | HPRT | 0.654 | TBP | 0.543 | HPRT | 7.64 | |
| UBC | 0.499 | UBC | 0.540 | ACTB | 0.738 | UBC | 0.811 | UBC | 9.15 | |
| All NFD/HFD (n = 32/32) |
GAPDH | 0.062 | HPRT | 0.278 | PPIA | 0.431 | RPLP0 | 0.403 | YWHAZ | 2.21 |
| YWHAZ | 0.090 | TBP | 0.278 | YWHAZ | 0.512 | YWHAZ | 0.460 | RPLP0 | 2.63 | |
| TBP | 0.108 | YWHAZ | 0.318 | RPLP0 | 0.557 | ACTB | 0.462 | GAPDH | 3.64 | |
| RPLP0 | 0.117 | RPLP0 | 0.331 | 18S | 0.559 | B2M | 0.490 | HPRT | 3.66 | |
| HPRT | 0.126 | GAPDH | 0.338 | TBP | 0.592 | GAPDH | 0.494 | TBP | 3.81 | |
| PPIA | 0.136 | ACTB | 0.359 | HPRT | 0.594 | HPRT | 0.499 | PPIA | 4.56 | |
| B2M | 0.144 | B2M | 0.387 | GAPDH | 0.615 | TBP | 0.502 | ACTB | 6.00 | |
| ACTB | 0.150 | 18S | 0.410 | B2M | 0.630 | PPIA | 0.506 | B2M | 6.29 | |
| 18S | 0.279 | PPIA | 0.433 | ACTB | 0.670 | 18S | 0.593 | 18S | 7.14 | |
| UBC | 0.419 | UBC | 0.609 | UBC | 1.289 | UBC | 1.101 | UBC | 10.00 | |
NFD, normal-fat diet; HFD, high-fat diet; All, all four points (4 w, 8 w, 12 w, 16 w); SD, standard deviation. Gene names indicated in bold are the most appropriate selection of 4 reference genes for all comparisons.