Table 2.
Analysis of reference gene expression variability in the epididymal fat 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) |
RPLP0 | 0.135 | PPIA | 0.244 | PPIA | 0.260 | YWHAZ | 0.429 | PPIA | 1.57 |
| PPIA | 0.139 | YWHAZ | 0.244 | 18S | 0.286 | RPLP0 | 0.430 | YWHAZ | 2.34 | |
| ACTB | 0.154 | 18S | 0.259 | YWHAZ | 0.305 | PPIA | 0.430 | RPLP0 | 2.66 | |
| B2M | 0.168 | ACTB | 0.298 | ACTB | 0.321 | ACTB | 0.467 | 18S | 3.66 | |
| YWHAZ | 0.183 | RPLP0 | 0.311 | RPLP0 | 0.366 | 18S | 0.509 | ACTB | 3.72 | |
| 18S | 0.233 | B2M | 0.333 | B2M | 0.419 | B2M | 0.514 | B2M | 5.42 | |
| HPRT | 0.308 | TBP | 0.381 | TBP | 0.466 | GAPDH | 0.562 | TBP | 7.48 | |
| TBP | 0.372 | GAPDH | 0.409 | GAPDH | 0.500 | TBP | 0.566 | GAPDH | 7.97 | |
| GAPDH | 0.438 | HPRT | 0.477 | HPRT | 0.530 | HPRT | 0.667 | HPRT | 8.45 | |
| UBC | 0.452 | UBC | 0.575 | UBC | 0.767 | UBC | 0.882 | UBC | 10.00 | |
| 8 w NFD/HFD (n = 8/8) |
B2M | 0.074 | RPLP0 | 0.172 | YWHAZ | 0.163 | RPLP0 | 0.309 | RPLP0 | 2.00 |
| RPLP0 | 0.090 | HPRT | 0.172 | 18S | 0.219 | HPRT | 0.321 | HPRT | 2.99 | |
| PPIA | 0.140 | B2M | 0.180 | TBP | 0.221 | PPIA | 0.335 | B2M | 3.03 | |
| HPRT | 0.149 | TBP | 0.202 | PPIA | 0.228 | B2M | 0.344 | PPIA | 3.83 | |
| TBP | 0.199 | ACTB | 0.223 | HPRT | 0.230 | ACTB | 0.345 | YWHAZ | 4.14 | |
| YWHAZ | 0.245 | PPIA | 0.236 | ACTB | 0.263 | TBP | 0.380 | TBP | 4.36 | |
| ACTB | 0.247 | YWHAZ | 0.247 | B2M | 0.265 | YWHAZ | 0.401 | ACTB | 5.69 | |
| GAPDH | 0.274 | 18S | 0.275 | RPLP0 | 0.287 | GAPDH | 0.421 | 18S | 6.00 | |
| 18S | 0.420 | GAPDH | 0.305 | GAPDH | 0.381 | 18S | 0.495 | GAPDH | 8.49 | |
| UBC | 0.942 | UBC | 0.472 | UBC | 1.030 | UBC | 0.985 | UBC | 10.00 | |
| 12 w NFD/HFD (n = 8/8) |
PPIA | 0.065 | RPLP0 | 0.168 | B2M | 0.176 | PPIA | 0.313 | RPLP0 | 1.68 |
| RPLP0 | 0.083 | B2M | 0.168 | RPLP0 | 0.212 | RPLP0 | 0.321 | PPIA | 1.86 | |
| B2M | 0.095 | PPIA | 0.198 | 18S | 0.213 | B2M | 0.326 | B2M | 2.06 | |
| YWHAZ | 0.155 | YWHAZ | 0.220 | PPIA | 0.275 | YWHAZ | 0.379 | YWHAZ | 4.43 | |
| HPRT | 0.188 | HPRT | 0.250 | GAPDH | 0.281 | GAPDH | 0.382 | 18S | 5.42 | |
| 18S | 0.208 | 18S | 0.272 | YWHAZ | 0.306 | ACTB | 0.385 | HPRT | 5.92 | |
| GAPDH | 0.267 | GAPDH | 0.297 | HPRT | 0.330 | HPRT | 0.405 | GAPDH | 5.92 | |
| ACTB | 0.282 | TBP | 0.313 | ACTB | 0.343 | 18S | 0.416 | ACTB | 7.67 | |
| TBP | 0.321 | ACTB | 0.328 | TBP | 0.358 | TBP | 0.451 | TBP | 8.74 | |
| UBC | 0.376 | UBC | 0.445 | UBC | 0.636 | UBC | 0.788 | UBC | 10.00 | |
| 16 w NFD/HFD (n = 8/8) |
RPLP0 | 0.083 | PPIA | 0.152 | TBP | 0.146 | PPIA | 0.323 | RPLP0 | 2.00 |
| HPRT | 0.098 | HPRT | 0.152 | RPLP0 | 0.230 | RPLP0 | 0.351 | PPIA | 2.11 | |
| B2M | 0.112 | B2M | 0.194 | GAPDH | 0.253 | YWHAZ | 0.356 | HPRT | 3.36 | |
| YWHAZ | 0.125 | RPLP0 | 0.232 | PPIA | 0.282 | HPRT | 0.364 | YWHAZ | 4.16 | |
| PPIA | 0.133 | YWHAZ | 0.244 | YWHAZ | 0.287 | B2M | 0.365 | B2M | 4.21 | |
| 18S | 0.141 | TBP | 0.274 | 18S | 0.298 | GAPDH | 0.402 | TBP | 4.28 | |
| TBP | 0.221 | GAPDH | 0.291 | B2M | 0.350 | 18S | 0.407 | GAPDH | 5.63 | |
| GAPDH | 0.285 | 18S | 0.303 | HPRT | 0.365 | TBP | 0.408 | 18S | 6.70 | |
| UBC | 0.330 | ACTB | 0.342 | ACTB | 0.567 | ACTB | 0.491 | ACTB | 9.24 | |
| ACTB | 0.384 | UBC | 0.439 | UBC | 0.662 | UBC | 0.817 | UBC | 9.74 | |
| All NFD/HFD (n = 32/32) |
PPIA | 0.041 | B2M | 0.286 | PPIA | 0.293 | RPLP0 | 0.488 | PPIA | 2.14 |
| RPLP0 | 0.047 | ACTB | 0.286 | YWHAZ | 0.324 | YWHAZ | 0.505 | RPLP0 | 2.38 | |
| YWHAZ | 0.093 | 18S | 0.343 | HPRT | 0.408 | PPIA | 0.542 | YWHAZ | 2.91 | |
| B2M | 0.111 | RPLP0 | 0.358 | RPLP0 | 0.527 | ACTB | 0.563 | B2M | 3.83 | |
| HPRT | 0.160 | TBP | 0.383 | TBP | 0.553 | HPRT | 0.568 | HPRT | 5.10 | |
| TBP | 0.213 | YWHAZ | 0.407 | 18S | 0.561 | B2M | 0.598 | ACTB | 5.63 | |
| 18S | 0.220 | PPIA | 0.421 | ACTB | 0.587 | GAPDH | 0.599 | 18S | 5.80 | |
| ACTB | 0.231 | GAPDH | 0.439 | GAPDH | 0.619 | TBP | 0.611 | TBP | 5.89 | |
| GAPDH | 0.291 | HPRT | 0.465 | B2M | 0.621 | 18S | 0.623 | GAPDH | 7.97 | |
| UBC | 0.372 | UBC | 0.742 | UBC | 1.366 | UBC | 1.508 | 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.