0.1

0.15

0.2

0.25

0.3

0 3 6 9 12 15 18 21 24 27 30 33 36 39 42 45 48 51 54 57 60 63 66 69 72 75 78 81 84 87 90

Absorbance@

590nm

Time(mins)

Glutamate

Malate

Isocitrate

Lactate

Co-culture PDAC tumor cells + CAFs respond to combination therapy of APX inhibition and metabolic inhibitor, CPI-613 (25 µM)

Arrows indicate treatment times.

0.00

0.20

0.40

0.60

0.80

1.00

1.20

1.40

1.60

1.80

4 8 12 14

Tumor Intensity Pa03C+CAF (n=3)

Ctrl CPI-613 APX3330 Combo

Days post plating

Fold

cha

nge

com

pare

d to

unt

reat

ed c

ontro

l

0.00

0.20

0.40

0.60

0.80

1.00

1.20

1.40

1.60

1.80

4 8 12 14

Tumor Intensity Pa03C+CAF (n=3)

Ctrl CPI-613 APX2009 Combo

APX3330=35 µM

Days post plating

APX2009=5 µM

-0.20

0.00

0.20

0.40

0.60

0.80

1.00

1.20

1.40

1.60

1.80

4 8 12 14

Tumor Intensity Pa03C+CAF (n=3)

Ctrl CPI-613 APX2014 Combo

Days post plating

APX2014=2.5 µM

Ctrl CPI-613 APX Combo

Co-culture PDAC tumor cells + CAFs respond to combination therapy of APX inhibition and metabolic inhibitor, CPI-613 (25 µM)

Arrows indicate treatment times.

0.00

0.20

0.40

0.60

0.80

1.00

1.20

1.40

1.60

1.80

4 8 12 14

Tumor Intensity Pa03C+CAF (n=3)

Ctrl CPI-613 APX3330 Combo

Days post plating

Fold

cha

nge

com

pare

d to

unt

reat

ed c

ontro

l

0.00

0.20

0.40

0.60

0.80

1.00

1.20

1.40

1.60

1.80

4 8 12 14

Tumor Intensity Pa03C+CAF (n=3)

Ctrl CPI-613 APX2009 Combo

APX3330=35 µM

Days post plating

APX2009=5 µM

-0.20

0.00

0.20

0.40

0.60

0.80

1.00

1.20

1.40

1.60

1.80

4 8 12 14

Tumor Intensity Pa03C+CAF (n=3)

Ctrl CPI-613 APX2014 Combo

Days post plating

APX2014=2.5 µM

Ctrl CPI-613 APX Combo

0

0.2

0.4

0.6

0.8

1

1.2

Day4 Day8 Day12

FoldCha

ngecompa

redtoSCR

G SCRG

SCRLowGSiGSiLowG

ScRNA-seq data of APE1 knockdown in normoxia and hypoxia

ScRNA-seq Differential Gene Expression Analysis

APEX1 Expression

Normoxia Hypoxia SCR SCR siAPE1 siAPE1

Fig 5: Validation of scRNA-seq in hypoxia. Expression of various Mitochondrial genes in normoxia and hypoxia, genes assessed via qRT-PCR in Pa03C cells. 3 independent experiments, showing average fold change +/- SD. *p