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A B
score Enrichment plot：NF⁃κB signaling pathway U251S N3S level 4 ** ** Vector
0.7
0.6
0.5
IBSP
Enrichment 0.4 0 p⁃p65 Vector IBSP Vector IBSP 60 kDa 3 2
0.3
0.2
0.1
p65 60 kDa Relative p⁃p65/p65 1
20
10 GAPDH 35 kDa 0
0 U251SN3rd N3T3rd
-10
0 5 000 10 000 15 000
Rank in ordered dataset sh⁃ctrl
1.5 sh⁃IBSP
level ** ** ** **
C U87 LN229 U251T3rd N3T3rd
sh⁃ctrl sh⁃IBSP sh⁃ctrl sh⁃IBSP sh⁃ctrl sh⁃IBSP sh⁃ctrl sh⁃IBSP 1.0

p⁃p65 60 kDa Relative p⁃p65/p65 0.5

p65 60 kDa
0
GAPDH 60 kDa U87 LN229 U251T3rd N3T3rd
D
IBSP SNAI1 TWIST1 CDH2 MYC U87 Vector
0.5 E ** IBSP
0.4 6
0.3 *** *** * * **
0.265 0.170 0.097 0.109 IBSP ** **
0.2
0.1 4
0 Relative expression
6.5
6.0
5.5 2
5.0 0.214 *** -0.245 *** 0.211 *** SNAI1
4.5
4.0
3.5 0
11 SNAI1 TWIST1 CDH2 MYC
9
7 -0.062 0.104 * TWIST1 Vector
5 IBSP
3 ** U251S **
10 6 ** **
8 -0.203 *** CDH2
6 Relative expression 4
13
11 2
9 MYC
7 0
5 SNAI1 TWIST1 CDH2 MYC
5 7 9 3.5 4.0 4.5 5.0 5.5 6.0 6.5 5 7 9 11 6 8 10 5 7 9 11 13
F U87 sh⁃ctrl U251T3rd sh⁃ctrl
1.5 1.5
sh⁃IBSP ** ** ** ** sh⁃IBSP
Relative expression 1.0 Relative expression 1.0
**
**
**
**
0.5
0.5
0 0
SNAI1 TWIST1 CDH2 MYC SNAI1 TWIST1 CDH2 MYC
A：The gene set enrichment analysis（GSEA）of IBSP in the TCGA dataset. B：p65 phosphorylation levels in control or IBSP overexpressed GBM
cells were measured by Western blot. C：p65 phosphorylation levels in control or IBSP depled GBM cells were measured by Western blot. D：Correlations
between IBSP and NF⁃κB targets were analyzed using TCGA dataset. E：mRNA levels of NF⁃κB targets in control or IBSP overexpressed cells were measured
**
***
*
by qRT⁃PCR. F：mRNA levels of NF⁃κB targets in control or IBSP depleted cells were measured by qRT⁃PCR. P < 0.05，P < 0.01， P < 0.001（n=3）.
图5 IBSP激活NF⁃κB信号通路
Figure 5 IBSP activated NF⁃κB signaling pathway