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Table 6 Constitutively active ALK1 regulated gene expression in primary HUVECs and the HUVEC cell line ECRF24. ECRF24 cells were infected with an MOI of ~50 with the recombinant adenovirus AdLacZ, containing the LacZ gene, or AdALK1QD, respectively. Primary HUVECs were infected with an MOI of ~50 with the empty AdCMV3 adenovirus or AdALK1QD, respectively. Gene expression was assessed after 24 hours by qRT-PCR. Non-infected cells served as reference. qRT-PCR based relative gene expression was calculated using the comparative ΔΔCT method. RQ represents the relative expression level (fold change) compared to the reference sample. RQMax/Min are the relative maximum/minimum expression levels of the mean expression level RQ. The right column titled HMEC-1 shows in comparison the mean RQ level of three independent experiments with HMEC-1 cells infected with AdALK1QD (see Table 4). Genes with RQ values in bold are considered to be up-regulated.

From: ALK1 signalling analysis identifies angiogenesis related genes and reveals disparity between TGF-β and constitutively active receptor induced gene expression

 

ECRF24

primary HUVECs

HMEC-1

Gene [GenBank number]

AdLacZ RQ (RQMax/Min)

AdALK1QDRQ (RQMax/Min)

AdCMV3 RQ (RQMax/Min)

AdALK1QDRQ (RQMax/Min)

AdALK1QDRQ

GADD153 [S40706]

0.95 (1.07/0.85)

4.21 (5.27/3.37)

1.17 (1.27/1.07)

7.53 (12.05/4.70)

4.55

IL-8 [M28130]

1.13 (1.01/1.27)

11.55 (13.60/9.81)

1.52 (2.55/0.90)

4.37 (5.57/3.44)

1056.07

SMAD6 [U59914]

0.82 (0.90/0.75)

5.65 (6.59/4.84)

1.30 (1.44/1.17)

38.89 (44.26/34.18)

14.56

COL5A1 [D90279]

1.04 (1.15/0.94)

1.68 (1.94/1.45)

1.00 (1.10/0.92)

3.47 (4.06/2.96)

2.28

HEF1 [L43821]

0.89 (0.98/0.81)

3.88 (4.55/3.31)

0.61 (0.66/0.56)

1.18 (1.75/0.80)

3.93

TLR4 [U93091]

0.95 (1.06/0.85)

1.59 (1.82/1.39)

1.02 (1.12/0.93)

4.77 (5.70/3.99)

3.98

ET-1 [J05008]

0.64 (0.70/0.59)

0.73 (0.86/0.61)

1.07 (1.16/0.99)

2.21 (2.62/1.87)

3.87

CARP [X83703]

0.61 (0.74/0.50)

0.79 (1.08/0.58)

0.83 (1.00/0.69)

1.94 (2.82/1.33)

4.29

HPTPη [D37781]

1.69 (1.90/1.51)

1.54 (2.75/0.86)

#

#

2.08

ID1 [S78825]

0.70 (0.78/0.63)

2.06 (2.45/1.73)

1.40 (1.79/1.09)

4.04 (6.63/2.46)

6.56

BMP6 [M60315]

0.64 (0.70/0.57)

0.72 (0.82/0.63)

0.91 (1.15/0.73)

1.61 (2.04/1.27)

4.26

ALK5 [L11695]

0.68 (0.78/0.59)

0.95 (1.25/0.71)

0.41 (0.44/0.38)

nd

1.36

TβRII [M85079]

1.18 (1.31/1.06)

0.77 (0.91/0.65)

0.96 (1.04/0.89)

1.38 (1.65/1.16)

1.26

Endoglin [J05481]

0.83 (0.93/0.74)

1.19 (1.60/0.89)

1.02 (1.10/0.95)

3.05 (4.07/2.29)

2.36

c-myc [V00568]

0.85 (1.03/0.70)

0.78 (0.90/0.67)

1.00 (1.17/0.86)

1.05 (1.64/0.67)

1.45

ALK1 [L17075] (ABI assay)

0.95 (1.09/0.83)

0.93 (1.11/0.77)

0.86 (0.93/0.79)

1.44 (2.28/0.91)

1.40

ALK1 [L17075] (own assay)

0.50

597.72*

0.55

197.18*

5079.92*

  1. nd, not detected after 40 cycles; #, not detected after 40 cycles and not detected in the reference sample; *, ectopic expression
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