Table 1

Percentage of apoptotic cells: comparison of results obtained with different methods in HT 1376 cells

UNTREATED CULTURES


METHOD

CULTURE TIME


6 hrs

16 hrs

24 hrs

48 hrs

72 hrs

96 hrs

120 hrs


Cell Cycle

HP

G0/1

S

G2/M

HP

G0/1

S

G2/M

HP

G0/1

S

G2/M

HP

G0/1

S

G2/M

HP

G0/1

S

G2/M

HP

G0/1

S

G2/M

HP

G0/1

S

G2/M

(PI staining)

1.0

42.6

37.4

20.0

1.0

44.0

35.4

20.6

2.0

47.9

30.5

21.6

2.6

60.5

31.0

8.5

2.1

64.5

25.7

9.8

3.3

70.1

19.3

10.6

5.6

78.1

12.3

9.6


6 hrs

16 hrs

24 hrs

48 hrs

72 hrs

96 hrs


JC-1

1.5

2.0

3.5

2.5

4.6

5.1

Low FSC area

4.4

5.6

8.9

11.8

10.8

30.7

Low FSC/SSC area

1.7

0.4

0.8

2.3

3.0

4.5

MAA+PI staining

E

L

N

E

L

N

E

L

N

E

L

N

E

L

N

E

L

N

N.E.

N.E.

N.E.

N.E.

N.E.

N.E.

0.5

0.6

5.4

1.5

1.3

7.2

3.1

3.4

10.6

5.0

5.8

10.8

AnnV+PI staining

E

L

N

E

L

N

E

L

N

E

L

N

E

L

N

E

L

N

N.E.

N.E.

N.E.

N.E.

N.E.

N.E.

2.9

0.7

4.5

2.0

3.2

5.1

1.2

4.7

7.3

3.1

5.9

8.2

TUNEL

N.E.

N.E.

0.5

0.8

1.6

1.5


DOCETAXEL-TREATED CULTURES


METHOD

CULTURE TIME


6 hrs

16 hrs

24 hrs

48 hrs

72 hrs

96 hrs

120 hrs


Cell Cycle

HP

G0/1

S

G2/M

HP

G0/1

S

G2/M

HP

G0/1

S

G2/M

HP

G0/1

S

G2/M

HP

G0/1

S

G2/M

HP

G0/1

S

G2/M

HP

G0/1

S

G2/M

(PI staining)

3.1

30.9

27.5

41.6

4.9

25.7

29.0

45.3

5.5

3.1

9.1

87.8

35.5

5.9

12.3

81.8

33.3

19.8

7.9

72.3

39.0

31.4

10.9

57.7

53.5

57.1

23.4

19.5


6 hrs

16 hrs

24 hrs

48 hrs

72 hrs

96 hrs


JC-1

3.0

10.4

24.5

53.6

60.0

91.3

Low FSC area

5.4

7.4

6.6

2.2

2.3

3.3

Low FSC/SSC area

6.5

8.5

24.2

29.6

32.5

33.6

MAA+PI staining

E

L

N

E

L

N

E

L

N

E

L

N

E

L

N

E

L

N

N.E.

N.E.

N.E.

N.E.

N.E.

N.E.

0.5

0.7

9.9

28.1

25.1

12.5

19.0

39.6

12.9

21.2

42.7

15.6

AnnV+PI staining

E

L

N

E

L

N

E

L

N

E

L

N

E

L

N

E

L

N

N.E.

N.E.

N.E.

N.E.

N.E.

N.E.

6.5

0.9

5.1

14.7

15.9

7.5

27.7

43.4

8.3

21.7

56.5

9.9

TUNEL

N.E.

N.E.

4.7

9.3

28.6

70.4


Top table: untreated (control) HT1376 bladder carcinoma cultures. Bottom table: HT1376 bladder carcinoma cultures were treated for 1 hour with 3 μg/mL docetaxel, after which the drug was washed out and the cells were further cultured. Figures are percentages of cells with the indicated characteristics. Data are the average of 3 independent experiments, with errors below 10%. For details, see the Methods section. N.E. = Not Examined; HP = Hypodiploid Peak; JC-1 = cells undergoing Δψm transition; E = early apoptosis, i.e. MAA+/PI- or AnnV+/PI-; L = late apoptosis, i.e. MAA+/PI+ or AnnV+/PI+; N = necrosis, i.e. MAA-/PI+ or AnnV-/PI+

Fabbri et al. BMC Cell Biology 2006 7:6   doi:10.1186/1471-2121-7-6

Open Data