1. AE1/AE3 Rabbit Normal IgG Supplementary Figure S1 White bar = 5 mm

2. KIF11 KIF23 NUF2 CDCA5 PLK1 CASC5 MAGEA2 NDC80 Supplementary Figure S2
KIF11 relative expression
KIF23 relative expression
NUF2
CDCA5
NUF2 relative expression
CDCA5 relative expression
PLK1
CASC5
PLK1 relative expression
CASC5 relative expression
MAGEA2
NDC80
MAGE-A2 relative expression
NDC80 relative expression

3. All histology Adenocarcinoma (ADC) P=0.7693 P=0.1104
Supplementary Figure S3
All histology
Adenocarcinoma (ADC)
1.0
1.0
KIF11-H (n = 156)
KIF11-H (n = 240)
0.8
0.8
0.6
0.6
Survival rate (x100%)
Survival rate (x100%)
KIF11-L (n =113)
KIF11-L (n =74)
0.4
0.4
P=0.7693
0.2
0.2
P=0.1104 2 4 6 8 10 12 2 4 6 8 10 12 14
(yrs)
(yrs)
KIF11-L (low-level KIF11: IHC score <0.25)
KIF11-H (high-level KIF11: IHC score ≥0.25) ○ □

4. mRNA knockdown Cell viability HPPS- siRNA- Scramble HPPS- siRNA- KIF11
Supplementary Figure S4
mRNA knockdown
Cell viability
HPPS-
siRNA-
Scramble
HPPS-
siRNA-
KIF11
HPPS-
siRNA-
Scramble
HPPS-
siRNA-
KIF11
No- Tf
No- Tf
Control
Control
H460SM cells: siRNA conc. = 600 nM

5. Change in tumor volume (%) Days after siRNAs injection
Supplementary Figure S5
Control (n=10)
Scramble (n=6)
Change in tumor volume (%)
KIF11 (n=6)
* P< (onSupplementary way ANOVA)
Days after siRNAs injection

6. H460 (Large Cell Carcinoma Cell line)
Supplementary Figure S6
H460 (Large Cell Carcinoma Cell line)
Cell viability (OD 490/630)
5.0 nM each
2.5 nM each
1.25 nM each
5.0 nM each
2.5 nM each
1.25 nM each
Cell viability (OD 490/630)
SBC5 (Small Cell Lung Cancer Cell line)
Before the in vivo study, we have conducted a cell viability assay with the combination of both KIF11- and PLK1 siRNA using the H460 cell line.
As you can see, we were able to confirm the synergetic effect of both KIF11- and PLK1 siRNA combination when we compare them with both Negative Control groups or siKIF11 or siPLK1 plus Negative Control groups at different siRNA concentrations. 
In particular, we can get 60% growth inhibition compared with Negative Control at the concentration of 5 nM each siRNAs.

7. Patient-specific siRNA-nanoparticle therapy
Supplementary Figure S7
Patient-specific siRNA-nanoparticle therapy
Biopsy sampling
from metastatic
LN using EBUS-TBNA
Copyright © Olympus Corporation
Identification of
target genes
expression pattern
using qRT-PCR
Symbol
Gene expression
Nanoparticle receptor
SCARB1
◎ highly-expressed
siRNA list
Candidate-1
KIF23
× no expressed
Candidate-2
KIF11
○ middle-expressed
Candidate-3
PLK1
Candidate-4
CDCA5
Candidate-5
NUF1
Candidate-6
CASC5
Candidate-7
Candidate-8
NDC80
siRNA-KIF11
Our goal is “Patient-specific nanoparticle-based lung cancer therapy.”
That is, at first, we will perform biopsy sampling from metastatic LNs using EBUS. Second, we will identify the gene expression pattern using RT-PCR analysis.
For example, if this tumor has high expression of receptor gene SB-R1, and also MAGEA2, and CDCA5 from the siRNA candidate genes list, we will be able to conjugate siRNAs against these genes to the nanoparticle. Then, we will use this combination therapy for the patients. Thus, our final aim is “Patient-derived, customized siRNA-nanoparticle therapy.”
nanoparticle with
siRNAs-KIF11
siRNA-PLK1
nanoparticle with
siRNAs-PLK1
Customized nanoparticle therapy
using patient-specific targeted siRNAs