Electronic Supplementary Material
Journal of Molecular Medicine, 2010
hSMG-1 is a Granzyme B-associated stress responsive protein kinase
F Meslin1, A Hamai1, B Mlecnik2, F Rosselli3, C Richon4, A Jalil1, G Wemhoff5, J Thiery1,6, J Galon2,
and S Chouaib 1*
1
Institut National de la Santé et de la Recherche Médicale (INSERM) U753; Laboratoire
d’immunologie des tumeurs humaines: interaction effecteurs cytotoxiques - système tumoral.
Institut Gustave Roussy PR1 and IFR 54, 94805 Villejuif Cedex, France
2
Institut National de la Santé et de la Recherche Médicale (INSERM) U872 ; centre de recherche des
Cordeliers ; 75006 Paris, France
3
Centre National de la Recherche Scientifique (CNRS) FRE2939, Laboratoire de stabilité génétique et
oncogenèse. Institut Gustave Roussy PR2 and IFR 54, 94805 Villejuif Cedex, France
4
Functional Genomic Unit, Institut Gustave Roussy PR2 and IFR 54, 94805 Villejuif Cedex, France
5
Functional Genomics, Sigma-Aldrich, 2909 Laclede Ave., St. Louis, MO 63103, USA.
6
Immune Disease Institute, Harvard Medical School, 200 Longwood Avenue, BOSTON, MA 02115
USA
*
Corresponding author: Dr. S. Chouaib, U753 INSERM, Institut Gustave Roussy, F-94805 Villejuif
Cedex,
FRANCE,
E-mail: [email protected]
Phone:
(33)
142114547,
Fax:
(33)
142115288,
Figure S1: Knockdown of CAD expression is sufficient to decrease the GrBmediated lysis in T1 melanoma tumor cells
Knockdown of CAD and BID expression in T1 cells was performed by RNA
interference and specific inhibition of caspase 3 was achieved by pre-incubation T1
cells with the chemical compound Z-DEVD-FMK (50μM) for 16h. For irrelevant
siRNA, T1 cells were pre-incubated with a negative control siRNA (LUC) targeting
luciferase. Cells were incubated with SLO alone or in combination with recombinant
human GrB for the indicated times. Analysis of GrB-induced apoptosis on T1 cells
was assessed by flow cytometry using Dioc6(3) and propidium iodide staining.
Figure S2: DNA damage response gene expression profiling of T1 cells
following CTL clone treatment
Cluster cDNA array analyses of T1 cells treated with CTL clone LT12 for 45min or
180min. Each column, a single cDNA chip experiment T1 treated with CTL clone
LT12 (180min (left), 45min (right)) compared with T1 cells.
Table 1 : Selected differentially expressed genes detected by cDNA microarray
analysis in T1 cells following GrB treatment (15min or 45min)
The changes for all of the genes are statistically significant. Log ratio expression of
selected differentially expressed genes detected by cDNA array analysis are
indicated.
Table 2 : Selected differentially expressed genes detected by cDNA microarray
analysis in T1 cells following CTL clone treatment (45min or 180min)
The changes for all of the genes are statistically significant. Log ratio expression of
selected differentially expressed genes detected by cDNA array analysis are
indicated.
Figure S2 (Supplemental information)
Figure S2: DNA damage response gene expression profiling of T1 cells following CTL clone treatment
Cluster cDNA array analyses of T1 cells treated with CTL clone LT12 for 45min or 180min. Each column, a
single cDNA chip experiment T1 treated with CTL clone LT12 (180min (left), 45min (right)) compared with T1
cells.
DNA damage pathway