Supplementary Figure 1: Silibinin has no considerable radiosensitizer effect on radioresponsive prostate cancer 22RV1 cells. (A) Effect of IR and SB on 22RV1 cell proliferation after 48 h of treatment. Exponentially growing 22RV1 cells were treated either with 2.5 or 5 Gy of IR and/or 25 μM SB. After 48 h of these treatments, cells were collected and total cells were counted using a hemocytometer. (B) Clonogenic assay was done with 22RV1 cells treated with 2.5 Gy IR alone or in combination with 25 μM SB. The number of colonies with greater than 50 cells were counted in each group at the end of 10 days. SB, silibinin; n.s., not significant. Control 25 μM SB 2.5 Gy I R + SB * * * B n.s. Control 25 μM SB 2.5 Gy 2.5 Gy + SB 5 Gy 5 Gy + SB * * * * * A n.s. n.s.
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Supplementary Figure 1: Silibinin has no considerable radiosensitizer effect on radioresponsive prostate cancer 22RV1 cells. (A) Effect of IR and SB on.
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Supplementary Figure 1: Silibinin has no considerable radiosensitizer effect on
radioresponsive prostate cancer 22RV1 cells. (A) Effect of IR and SB on 22RV1 cell
proliferation after 48 h of treatment. Exponentially growing 22RV1 cells were treated
either with 2.5 or 5 Gy of IR and/or 25 μM SB. After 48 h of these treatments, cells
were collected and total cells were counted using a hemocytometer. (B) Clonogenic
assay was done with 22RV1 cells treated with 2.5 Gy IR alone or in combination with
25 μM SB. The number of colonies with greater than 50 cells were counted in each
group at the end of 10 days. SB, silibinin; n.s., not significant.
Con
trol
25
μM S
B
2.5
Gy
IR +
SB
*
**
B
n.s.
Con
trol
25
μM S
B
2.5
Gy
2.5
Gy
+ SB
5 G
y5
Gy
+ SB
*
*
*
**
A
n.s.
n.s.
Supplementary Figure 2. Silibinin potentiates and prolongs G2/M arrest by IR
exposure in DU145 cells. For cell cycle analysis, DU145 cells were exposed to IR (5
Gy) with or without silibinin (25 SB. After 24 and 48 h of treatments, cells were
processed for saponin/PI staining. Representative histogram showing a prominent
G2/M arrest in combination at 24 and 48 h in DU145 cells. The quantitative data is
shown as figure 2A.
Control 25 M SB 5 Gy IR IR+SB
24 h
48 h
IR+S
B
Con
trol
S
B IR
Con
trol
PC-3DU145
SurvivinPCNA
BIR
+SB
S
B IR
IR+S
B
Con
trol
S
B IR
Con
trol
PC-3DU145
IR+S
B
S
B IR
A
Cyclin B1
Cdc-2
IR+S
B
Con
trol
SB IR
GAPDH
Cdc25C
DU145
Supplementary Figure 3. Silibinin decreases the molecules regulating G2/M phase
transition in PCa cells, but not in HEK-293 cells in response to IR exposure, and
inhibits cell proliferation and survival. (A) mRNA expression levels of G2/M phase
cell cycle regulatory molecules after 48 h of treatment with IR (5 Gy) and /or SB (25
μM) in DU145 and HEK-293 cells. GADPH was used as loading control. (B)
Densitometric quantitation data showing the fold change in the protein levels of PCNA
and survivin (from figure 2D) after indicated treatments in DU145 and PC-3 cells.