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Table 1 Regulation of 38 barrier targets on the mRNA level in hCMEC/D3 samples, which were treated either with 5% FCS, 0.25% FCS or 0.25% FCS plus 100 nM hydrocortisone (HC) in EBM-2 medium prior OGD treatment

From: The pivotal role of micro-environmental cells in a human blood–brain barrier in vitro model of cerebral ischemia: functional and transcriptomic analysis

Day 5 on 6

5% EBM-2

0.25% EBM-2

0.25% EBM-2 + HC

5% EBM-2

0.25% EBM-2

0.25% EBM-2 + HC

5% EBM-2

0.25% EBM-2

0.25% EBM-2 + HC

5 h treatment

5% EBM-2

0.25% EBM-2

0.25% EBM-2 + HC

DMEM + Glu

DMEM + Glu

DMEM + Glu

DMEM–Glu

DMEM–Glu

DMEM–Glu

Normoxia/OGD

N

N

N

N

N

N

OGD

OGD

OGD

Claudin-1

0.63 ± 0.10*

0.62 ± 0.07*

0.46 ± 0.07*

0.94 ± 0.10

1.00 ± 0.00

0.92 ± 0.15

0.83 ± 0.07*§

1.52 ± 0.10*

0.77 ± 0.06*§

Claudin-3

1.25 ± 0.24

1.89 ± 0.88

1.83 ± 0.53

0.80 ± 0.14

1.00 ± 0.00

1.38 ± 0.32

0.58 ± 0.28

0.44 ± 0.11*

1.86 ± 0.93

Claudin-4

1.28 ± 0.24

1.49 ± 0.31

1.97 ± 0.42*

0.85 ± 0.08

1.00 ± 0.00

1.19 ± 0.17

0.66 ± 0.22

0.59 ± 0.07*

1.92 ± 0.89

Claudin-5

1.17 ± 0.26

1.51 ± 0.47

3.00 ± 0.72*

1.03 ± 0.11

1.00 ± 0.00

1.42 ± 0.30

1.31 ± 0.32

1.35 ± 0.14*

3.98 ± 1.40

Claudin-6

1.48 ± 0.49

1.93 ± 0.86

2.54 ± 0.98

0.97 ± 0.19

1.00 ± 0.00

1.43 ± 0.40

1.39 ± 0.70

1.07 ± 0.18

5.66 ± 3.09

Claudin-7

1.41 ± 0.22

1.14 ± 0.08

1.21 ± 0.08*

1.28 ± 0.15

1.00 ± 0.00

0.97 ± 0.11

0.96 ± 0.09

0.88 ± 0.10

1.08 ± 0.22

Claudin-8

7.28 ± 4.89

7.41 ± 4.06

18.02 ± 12.19

2.09 ± 1.76

1.00 ± 0.00

3.13 ± 1.81

10.05 ± 8.27

2.40 ± 0.95

53.64 ± 35.49

Claudin-9

3.25 ± 1.48

3.93 ± 2.04

12.19 ± 8.17

1.20 ± 0.60

1.00 ± 0.00

1.82 ± 0.63

5.44 ± 4.16

0.80 ± 0.16

20.00 ± 13.21

Claudin-10 tva

3.91 ± 2.46

4.77 ± 2.16

9.09 ± 5.88

1.31 ± 1.02

1.00 ± 0.00

1.96 ± 1.11

5.58 ± 4.60

1.96 ± 0.74

29.76 ± 19.82

Claudin-10 tvb

14.02 ± 8.92

8.74 ± 5.73

26.92 ± 18.18

4.12 ± 3.57

1.00 ± 0.00

4.53 ± 2.51

11.80 ± 10.29

2.21 ± 0.79

61.32 ± 40.30

Claudin-11

1.02 ± 0.13

0.81 ± 0.05*

0.58 ± 0.06#*

1.28 ± 0.18

1.00 ± 0.00

0.78 ± 0.05*

0.66 ± 0.05*

0.82 ± 0.09

0.54 ± 0.06*§

Claudin-12 tv1

1.43 ± 0.12*

1.48 ± 0.18*

1.78 ± 0.14*

0.98 ± 0.08

1.00 ± 0.00

1.00 ± 0.10

0.83 ± 0.05*

1.00 ± 0.12

1.06 ± 0.09

Claudin-12 tv2

1.73 ± 0.21*

1.72 ± 0.19*

1.96 ± 0.08*

1.17 ± 0.13

1.00 ± 0.00

1.07 ± 0.13

1.27 ± 0.09*§

1.87 ± 0.19*

1.98 ± 0.28*

Claudin-12 tv3

1.17 ± 0.16

1.26 ± 0.13

1.31 ± 0.04*

1.14 ± 0.12

1.00 ± 0.00

0.95 ± 0.10

0.99 ± 0.08§

1.63 ± 0.14*

1.49 ± 0.24

Claudin-14

1.22 ± 0.14

0.77 ± 0.14

0.84 ± 0.13

1.01 ± 0.07

1.00 ± 0.00

1.16 ± 0.29

0.75 ± 0.08*

0.99 ± 0.16

0.52 ± 0.07*§

Claudin-15

1.01 ± 0.13

0.90 ± 0.12

1.77 ± 0.47

1.17 ± 0.07*

1.00 ± 0.00

1.41 ± 0.34

0.81 ± 0.12§

1.22 ± 0.14

1.88 ± 0.22*§

Claudin-16

1.81 ± 0.28*

1.62 ± 0.32

1.25 ± 0.20

0.99 ± 0.11

1.00 ± 0.00

1.07 ± 0.21

0.70 ± 0.05*§

1.08 ± 0.11

0.66 ± 0.12*§

Claudin-17

12.78 ± 8.63

11.44 ± 8.08

35.16 ± 24.62

4.33 ± 3.99

1.00 ± 0.00

5.63 ± 3.59

20.47 ± 18.10

2.14 ± 0.57

119.56 ± 83.89

Claudin-18 tva

1.16 ± 0.26

1.07 ± 0.23

2.97 ± 1.20

1.09 ± 0.13

1.00 ± 0.00

1.97 ± 0.70

0.23 ± 0.07*

0.31 ± 0.08*

0.88 ± 0.39

Claudin-18 tvb

1.53 ± 0.57

3.03 ± 1.25

4.07 ± 2.79

0.61 ± 0.27

1.00 ± 0.00

0.77 ± 0.25

1.89 ± 1.37

1.07 ± 0.33

8.96 ± 5.38

Claudin-20

1.24 ± 0.17

1.19 ± 0.27

2.04 ± 0.59

0.66 ± 0.13*

1.00 ± 0.00

1.77 ± 0.49

0.25 ± 0.10*

0.38 ± 0.12*

0.63 ± 0.22

Claudin-22

0.81 ± 0.11

0.85 ± 0.08

1.18 ± 0.16

0.99 ± 0.10

1.00 ± 0.00

1.14 ± 0.17

0.62 ± 0.12*

0.93 ± 0.14

1.57 ± 0.45

Claudin-23

0.82 ± 0.08*

0.99 ± 0.07

0.92 ± 0.21

0.80 ± 0.15

1.00 ± 0.00

0.70 ± 0.17

0.64 ± 0.15*

0.95 ± 0.16

1.72 ± 0.76

Claudin-24

0.87 ± 0.15

0.98 ± 0.28

1.51 ± 0.31

1.05 ± 0.13

1.00 ± 0.00

1.62 ± 0.36

0.57 ± 0.17*

0.81 ± 0.13

2.34 ± 0.98

Claudin-25

3.47 ± 1.94

8.17 ± 4.46

8.08 ± 5.39

1.04 ± 0.65

1.00 ± 0.00

1.77 ± 0,76

4.02 ± 2.93

3.39 ± 0.14

22.29 ± 14.18

ZO-1

0.80 ± 0.15

0.65 ± 0.06*

1.16 ± 0.42

1.20 ± 0.20

1.00 ± 0.00

1.78 ± 0.56

0.86 ± 0.11

1.53 ± 0.33

1.62 ± 0.33

ZO-2

0.81 ± 0.08*

0.93 ± 0.08

1.26 ± 0.17

0.99 ± 0.09

1.00 ± 0.00

1.16 ± 0,19

0.86 ± 0.08§

1.39 ± 0.14*

1.61 ± 0.21*

ZO-3

1.21 ± 0.24

2.12 ± 0.70

3.81 ± 0.96*

1.37 ± 0.39

1.00 ± 0.00

3.39 ± 0.72*

0.79 ± 0.21§

1.71 ± 0.30*

2.80 ± 0.42*

JAM-1

0.90 ± 0.07#

1.27 ± 0.11*

1.42 ± 0.08*

0.94 ± 0.12

1.00 ± 0.00

1.14 ± 0.18

0.56 ± 0.05*§

0.80 ± 0.07*

0.87 ± 0.12

JAM-2

0.69 ± 0.31

1.23 ± 0.24

1.92 ± 0.42*

1.45 ± 0.70

1.00 ± 0.00

0.84 ± 0.23

0.79 ± 0.13

1.30 ± 0.21

0.54 ± 0.13*§

JAM-3

0.91 ± 0.12

1.43 ± 0.39

1.13 ± 0.11

1.05 ± 0.13

1.00 ± 0.00

0.94 ± 0.07

0.84 ± 0.08

1.13 ± 0.11

1.42 ± 0.22

Occludin

0.89 ± 0.10

0.78 ± 0.06*

1.23 ± 0.29

1.00 ± 0.12

1.00 ± 0.00

1.67 ± 0.54

0.85 ± 0.10§

1.28 ± 0.15

1.23 ± 0.15

Tricellulin

1.00 ± 0.07

1.21 ± 0.11

2.56 ± 0.98

0.84 ± 0.09

1.00 ± 0.00

1.56 ± 0.40

0.74 ± 0.05*§

1.05 ± 0.07

1.27 ± 0.14

CDH5

0.81 ± 0.09

0.83 ± 0.03*

1.04 ± 0.26

0.94 ± 0.09

1.00 ± 0.00

1.11 ± 0.27

0.80 ± 0.05*§

1.23 ± 0.09*

1.05 ± 0.14

ABCB1 (PgP)

0.82 ± 0.11

0.79 ± 0.06*

1.81 ± 0.53

1.30 ± 0.16

1.00 ± 0.00

1.61 ± 0.50

1.01 ± 0.09

1.07 ± 0.13

1.41 ± 0.19

ABCC1 (MRP1)

1.28 ± 0.16

1.00 ± 0,10

2.31 ± 0.96

1.20 ± 0.10

1.00 ± 0.00

1.88 ± 0.71

1.13 ± 0.19

1.27 ± 0.09*

1.48 ± 0.16*

ABCC2 (MRP2)

1.22 ± 0.12#

0.94 ± 0.03*

1.45 ± 0.37

1.25 ± 0.14

1.00 ± 0.00

1.45 ± 0.60

0.99 ± 0.07

1.13 ± 0.12

1.04 ± 0.16

ABCC3 (MRP3)

0.91 ± 0.10

0.91 ± 0.06

2.19 ± 0.16#*

0.85 ± 0.09

1.00 ± 0.00

1.94 ± 0.41*

0.66 ± 0.07*

0.72 ± 0.04*

1.65 ± 0.28*§

ABCC4 (MRP4)

0.65 ± 0.06*

0.68 ± 0.02*

1.33 ± 0.30

1.01 ± 0.09

1.00 ± 0.00

1.32 ± 0.37

0.55 ± 0.05*

0.59 ± 0.06*

0.74 ± 0.12

ABCC5 (MRP5)

1.27 ± 0.21

1.14 ± 0.07

2.54 ± 0.90

1.22 ± 0.13

1.00 ± 0.00

1.52 ± 0.50

1.11 ± 0.11

1.45 ± 0.15*

1.88 ± 0.24*

ABCG2 (BCRP)

1.04 ± 0.10

1.14 ± 0.08

1.48 ± 0.16*

1.01 ± 0.10

1.00 ± 0.00

1.20 ± 0.26

1.04 ± 0.07

1.28 ± 0.15

1.49 ± 0.25

SLC2A1 (Glut1)

1.41 ± 0.16#*

0.85 ± 0.07*

0.85 ± 0.15

1.00 ± 0.10

1.00 ± 0.00

1.19 ± 0.25

2.62 ± 0.36*

3.63 ± 0.42*

4.09 ± 0.51*

VEGFa

2.93 ± 0.50*

1.26 ± 0.14

1.25 ± 0.36

1.06 ± 0.11

1.00 ± 0.00

1.29 ± 0.40

4.46 ± 0.62*

5.63 ± 0.49*

5.83 ± 0.57*

  1. Data were normalized on β-actin (n = 8 from four independent experiments). Claudin-2 und—19 were not detected, claudin-13 and -21 are not present in human, tv = transcript variant. Data are presented as mean ± SEM of the x-fold regulation related to hCMEC/D3 cells, which were cultivated from day 5 to 6 in EBM-2 with 0.25% FCS, their medium was changed on day 6 in DMEM plus glucose and treated for 5 h under normoxic conditions. Statistically significant regulations were marked with: *p < 0.05 versus 0.25% EBM-2 (day 5 to 6) and DMEM + Glu (5 h treatment) and N (normoxia); #p < 0.05 versus 0.25% EBM-2 (day 5 to 6) and 0.25% EBM-2 (5 h treatment) and N (normoxia); §p < 0.05 versus 0.25% EBM-2 (day 5 to 6) and DMEM—Glu (5 h treatment) and OGD (oxygen/glucose deprivation
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Fluids and Barriers of the CNS

ISSN: 2045-8118