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Table 4 Quantitative RT-PCR analysis of gene expression during rapid trophoblastic elongation for SAHH, HSC70, and OSU-T1-50.

From: Analysis and characterization of differential gene expression during rapid trophoblastic elongation in the pig using suppression subtractive hybridization

Transcript

Morphology

Average Target CT *

Average 18S rRNA CT *

ΔCT‡¶

ΔΔCT§

SAHH

Spherical

23.48 ± 0.23

17.47 ± 0.10

6.01 ± 0.21a

-0.20

 

Ovoid

23.62 ± 0.82

17.78 ± 0.24

5.84 ± 0.58a

-0.37

 

Tubular

23.20 ± 0.51

16.99 ± 0.14

6.21 ± 0.51a

0.00

 

Filamentous

20.72 ± 0.55

17.24 ± 0.12

3.47 ± 0.58b

-2.76

HSC70kD

Spherical

20.86 ± 0.28

17.47 ± 0.10

3.38 ± 0.25c

0.00

 

Ovoid

21.02 ± 0.80

17.78 ± 0.24

3.24 ± 0.57c

-0.15

 

Tubular

20.17 ± 0.52

16.99 ± 0.14

3.18 ± 0.48c

-0.21

 

Filamentous

17.35 ± 0.46

17.24 ± 0.12

0.11 ± 0.48d

-3.27

OSU-T1-50

Spherical

33.22 ± 0.48

17.47 ± 0.10

15.75 ± 0.64e

0.00

 

Ovoid

32.60 ± 1.01

17.78 ± 0.24

14.82 ± 1.22ef

-0.93

 

Tubular

29.21 ± 0.63

16.99 ± 0.14

12.22 ± 0.80f

-3.53

 

Filamentous

26.30 ± 0.57

17.24 ± 0.12

9.05 ± 0.80g

-6.69

  1. Quantitative RT-PCR analysis comparing gene expression across morphologies during rapid trophoblastic elongation for three of the transcripts identified using suppression subtractive hybridization: s-adenosylhomocysteine hydrolase, heat shock cognate 70 kDa, and an unknown transcript identified by its clone number, OSU-T1-50. * CT = Cycle Threshhold. Indicates cycle number in which amplification crosses the threshold set in the geometric portion of amplification curve. ‡ ΔCT = Target transcript CT – 18S ribosomal CT: Normalization of CT for target gene relative to ribosomal 18S RNA CT. ¶Statistical analysis of normalized expression levels between morphologies. Values with different superscripts for each of the target genes differ significantly: ab(P < 0.01), cd(P < 0.001), ef(P < 0.004),eg(P < 0.001), and fg(P < 0.01). §ΔΔCT = Mean ΔCT – highest mean ΔCT value: The mean value for the morphology with highest ΔCT (lowest expression levels for target) was used as a calibrator to set the baseline for comparing mean differences in the ΔCT values across all morphologies.