SNU423 SK-Hep1 HepG2 HA22T

AR

GAPDH

Supplemental Fig. S1 Endogenous AR expression in various HCC cell lines

Supplemental Fig. S2 Multiple E:T ratio for NK cells against HCC cells

Supplemental Fig. S3 Tunel assay for NK cells against SNU-423 cells

SNU423-Scr SNU423-shAR0

5

10

15**

Apop

tosi

s ra

te (f

old

incr

ease

)

SNU423-Scr SNU423-shAR

0 2 6 12 240

100

200

300

400

SK-pBABESK-AR3SK-AR7SNU423-ScrSNU423-shAR

*NS

*

*NS

**

***

*** ***

hours

IL-1

2 p3

5 (p

g/m

l)

Supplemental Fig. S4 IL-12p35 release of HCC cells with various AR

Supplemental Fig. S5 MAGNET V2.0 calculated correlation between IL12A and AR

Nontumor Peritumor Intratumor

Patient 22CD56

Supplemental Fig. S6 Distribution of NK cells in different region of HCC

Nontumor Paritumor Intratumor0

20

40

60 *

***

CD

56+

cells

per

fiel

d

SK-pBabe SK-AR70

20

40

60

80

100

120

-++++++

Spearman R= -0.329P value= 0.1165

n=12 n=12

Perc

enta

ge %

Vehicle Sorafenib0

20

40

60

80

100

120

-++++++

n=6 n=6 IL-12A

Perc

enta

ge %

Supplemental Fig. S9 Correlation analysis for animal tumor samples staining

0 4 6 80

100

200

300

400 **

*SK-Hep1

+sorafenib (umol/L)

IL-1

2 p3

5 (p

g/m

l)

Supplemental Fig. S7 IL-12p35 release induced by sorafenib treatment

0 4 6 80

100

200

300

400 ***

*SNU-423

+sorafenib (umol/L)

IL-1

2 p3

5 (p

g/m

l)

Supplemental Fig. S8 Ectopic AR in HCC cells interrupted sorafenib induced IL-12A

Supplemental figure 1. Different expression level of AR in HCC cells lines we applied. Supplemental figure 2. LDH cytotoxic assay performed with HA22T-AR (upper panel) or SK-AR (lower panel) cells. Multiple E:T ratios were compared. Data shown are mean±SEM. *** P< 0.001, ** P<0.01, * P<0.05.

Supplemental figure 3. TUNEL staining for second HCC cell line, SNU423, after adding NK-92MI cells. Left panel is representative TUNEL staining image and right panel is quantification result. *** P< 0.001, ** P<0.01, * P<0.05. Supplemental figure 4. Culture media was collected from HCC cells with different AR expression at different time points. Then we detected IL-12p35 in media with an ELISA kit. *** P< 0.001, ** P<0.01, * P<0.05.

Supplemental figure 5. Correlation analysis result generated from MAGNET V2.0 website by using TCGA database.

Supplemental figure 6. Representative image for CD56+ NK cells in nontumor, peritumor and intratumor tissue, and CD56+ cells quantification. *** P< 0.001, ** P<0.01, * P<0.05.

Supplemental figure 7. Culture media was collected from HCC cells treated with sorafenib for 24 hours. Then we detected IL-12p35 in media with an ELISA kit. *** P< 0.001, ** P<0.01, * P<0.05.

Supplemental figure 8. Ectopic AR was provided into HCC cells by lentivirus, then treated with 4uM sorafenib for further testing IL-12A expression with Western Blot.

Supplemental figure 9. Quantification results for staining IL-12A and AR in orthotopic HCC tumors from nude mice. Left panel is comparing AR(+) and AR(-). Right panel is showing the difference after sorafenib treatment.