Ren ONE58 NCI-H28

- + - + - + 5M SAHA

1M MG132

FLIPL

FLIPS

β-actin

10MZ-VAD

PARP

FLIPL

FLIPS

p18-caspase 8

β-actin

A

B

C

Time (hrs) 3 6 12 24 SAHA - + - + - + - +

3 6 12 24

- + - + - + - +

3 6 12 24

- + - + - + - +

Ren ONE58 NCI-H28

Fig. S2

Fig. S2 - The kinetics of FLIP down-regulation by SAHA. Ren, ONE58 and NCI-H28 cells were

treated with 5μM SAHA for 3, 6, 12 and 24 h. (A) qPCR analysis of total FLIP (FLIPL and

FLIPS) mRNA levels following SAHA treatment, normalised to GAPDH mRNA expression. (B)

Western blot analysis showing FLIPL and FLIPS down-regulation, caspase 8 processing and

PARP cleavage in each cell line. (C) Ren cells were pre-treated with 10 µM z-VAD-fmk (pan-

caspase inhibitor) or 1 µM MG132 (proteasome inhibitor) for 1 h, followed by treatment with 5

μM SAHA for 12 h. Immunoblot analysis shows that proteasome inhibition attenuates SAHA-

induced FLIP down-regulation, however caspase inhibition fails to prevent SAHA-induced FLIP

down-regulation.