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Table 1 The rank of candidate reference gene stability for monocultured OCCM-30 or SV-PDL, direct cell–cell culture, co-cultured OCCM-30 cells and co-cultured SV-PDL cells, overall ranks were calculated by the four algorithms application (geNorm, NormFinder, comparative ΔCq and BestKeeper, respectively)

From: Selection and validation of reference gene for RT-qPCR studies in co-culture system of mouse cementoblasts and periodontal ligament cells

Rank Methods   geNorm   NormFinder   Comparative ΔCq   BestKeeper
Ranking order Ranking sum Ranking order Stability value (M) Ranking order Stability value Ranking order Stability value Ranking order Stability value (r) SD
(± Cq)
CV
(% Cq)
Monocultured cementoblasts (OCCM-30)
 (1) RPL22 13   GUSB 0.032   TBP 0.065   RPL22 0.22   PPIB 0.750 0.470 2.278
 (2) PPIB 17   POLR2A 0.037   PPIB 0.065   RPLP0 0.26   YWHAZ 0.685 1.541 7.321
 (3) POLR2A 21   RPL22 0.042   β-actin 0.092   POLR2A 0.40   GAPDH 0.679 0.390 2.282
 (4) RPLP0 23   RPLP0 0.058   RPL22 0.136   GUSB 0.40   TBP 0.620 0.503 2.121
 (5) GUSB 23   UBC 0.063   RPL 0.212   UBC 0.53   RPL22 0.612 0.662 2.679
 (6) β-actin 24   β-actin 0.072   RPLP0 0.538   PPIB 0.54   β-actin 0.453 0.543 3.208
 (7) TBP 24   GAPDH 0.083   POLR2A 0.719   EEF1A1 0.59   RPL 0.172 0.204 0.790
 (8) UBC 27   PPIB 0.090   GUSB 0.831   TBP 0.59   UBC -0.038 0.489 1.756
 (9) GAPDH 32   YWHAZ 0.095   UBC 0.859   β-actin 0.64   POLR2A -0.115 0.197 0.875
 (10) YWHAZ 35   EEF1A1 0.113   EEF1A1 1.005   RPL 0.81   GUSB -0.282 0.331 1.393
 (11) EEF1A1 39   TBP 0.183   GAPDH 2.421   GAPDH 2.12   RPLP0 -0.428 0.152 0.988
 (12) RPL -      YWHAZ 2.461   YWHAZ 2.29   EEF1A1 -0.683 0.472 2.774
 (13) TUBB -         TUBB 13.85   TUBB -0.986 0.295 0.795
Monocultured periodontal ligament cells (SV-PDL)
 (1) GUSB 14   GUSB 0.003   EEF1A1 0.112   GUSB 0.18   TBP 0.928 0.447 1.766
 (2) RPLP0 19   GAPDH 0.004   GUSB 0.112   RPLP0 0.19   RPL22 0.921 0.762 2.951
 (3) RPL22 22   RPLP0 0.007   POLR2A 0.123   EEF1A1 0.19   UBC 0.916 0.660 2.360
 (4) TBP 22   β-actin 0.021   β-actin 0.124   POLR2A 0.24   YWHAZ 0.910 0.455 2.103
 (5) β-actin 22   PPIB 0.031   TBP 0.124   RPL22 0.26   β-actin 0.905 0.537 3.093
 (6) PPIB 26   RPL22 0.040   RPLP0 0.125   TUBB 0.31   GAPDH 0.868 0.518 2.894
 (7) POLR2A 27   UBC 0.045   PPIB 0.206   PPIB 0.51   PPIB 0.739 0.446 2.094
 (8) GAPDH 30   TBP 0.050   TUBB 0.282   TBP 0.53   RPLP0 0.440 0.126 0.818
 (9) TUBB 36   POLR2A 0.075   RPL22 0.475   β-actin 0.66   RPL 0.355 0.250 0.996
 (10) EEF1A1 38   TUBB 0.100   RPL 0.526   RPL 0.92   GUSB 0.328 0.138 0.561
 (11) YWHAZ 49   EEF1A1 0.127   GAPDH 5.241   GAPDH 4.98   POLR2A 0.169 0.226 0.989
 (12) UBC -   YWHAZ 0.850   YWHAZ 6.351   YWHAZ 5.99   TUBB 0.125 0.268 0.930
 (13) RPL -         UBC 10.46   EEF1A1 -0.292 0.168 0.967
Direct cell–cell contact cultured of OCCM-30 and SV-PDL
 (1) GUSB 7   POLR2A 0.086   GUSB 0.005   RPLP0 0.29   POLR2A 0.912 0.57 2.27
 (2) POLR2A 8   GUSB 0.089   POLR2A 0.007   GUSB 0.48   GUSB 0.889 0.62 2.36
 (3) RPLP0 14   RPL22 0.11   RPL22 0.019   PPIB 0.48   PPIB 0.872 1.07 4.83
 (4) RPL22 15   RPLP0 0.212   RPLP0 0.029   POLR2A 0.59   RPL22 0.784 0.62 2.22
 (5) PPIB 16   PPIB 0.399   PPIB 0.058   RPL22 1.09   RPLP0 0.575 0.31 2.00
Co-cultured OCCM-30 control
 (1) PPIB 8   PPIB 0.211   GUSB 0.006   RPLP0 0.31   RPL22 0.986 1.5 5.31
 (2) GUSB 10   POLR2A 0.224   PPIB 0.013   GUSB 1.00   PPIB 0.977 0.95 4.34
 (3) RPLP0 14   RPLP0 0.325   RPL22 0.028   PPIB 1.02   GUSB 0.977 0.94 3.46
 (4) POLR2A 14   GUSB 0.347   POLR2A 0.040   POLR2A 1.03   POLR2A 0.658 1.01 3.92
 (5) RPL22 14   RPL22 0.704   RPLP0 0.045   RPL22 1.59   RPLP0 0.220 0.26 1.65
Co-cultured OCCM-30 with 3 indicated time (n = 18)
 (1) PPIB 9   RPLP0 0.409   PPIB 0.016   RPLP0 0.63   PPIB 0.949 0.72 3.25
 (2) GUSB 9   GUSB 0.478   GUSB 0.021   POLR2A 0.74   GUSB 0.767 0.6 2.18
 (3) RPLP0 11   PPIB 0.515   POLR2A 0.027   GUSB 0.78   RPL22 0.712 0.98 3.54
 (4) POLR2A 14   POLR2A 0.565   RPL22 0.031   PPIB 0.78   RPLP0 0.500 0.49 2.9
 (5) RPL22 17   RPL22 0.840   RPLP0 0.035   RPL22 1.27   POLR2A 0.427 0.64 2.48
Co-cultured SV-PDL control
 (1) PPIB 9   RPLP0 0.308   PPIB 0.004   RPLP0 0.11   POLR2A 0.998 0.65 2.59
 (2) POLR2A 11   PPIB 0.309   RPL22 0.004   PPIB 0.52   RPL22 0.983 0.68 2.59
 (3) RPLP0 12   POLR2A 0.378   POLR2A 0.009   RPL22 0.62   GUSB 0.980 0.53 1.85
 (4) RPL22 12   GUSB 0.390   GUSB 0.011   POLR2A 0.74   PPIB 0.973 0.47 2.17
 (5) GUSB 16   RPL22 0.732   RPLP0 0.023   GUSB 0.78   RPLP0 0.700 0.09 0.61
Co-cultured SV-PDL with 3 indicated time (n = 18)
 (1) PPIB 8   POLR2A 0.161   PPIB 0.005   RPLP0 0.22   PPIB 0.980 0.31 1.44
 (2) POLR2A 10   GUSB 0.165   RPLP0 0.015   PPIB 0.42   POLR2A 0.823 0.5 1.98
 (3) RPLP0 11   RPLP0 0.214   GUSB 0.016   POLR2A 0.60   GUSB 0.774 0.54 2.04
 (4) GUSB 12   PPIB 0.218   POLR2A 0.017   GUSB 0.60   RPL22 0.746 0.57 2.02
 (5) RPL22 19   RPL22 0.422   RPL22 0.019   RPL22 0.74   RPLP0 0.646 0.19 1.21
  1. A higher rank denotes lower expression stability. Individual primer efficiency was taken in to account by the abbreviations of Cq (threshold cycle), SD (standard deviation), CV (coefficient of variation) and r (Pearson’s correlation coefficient). The genes are ordered form the highest to the lowest ranking