Supplementary Figure 1
A
B
GPR81
shControl
shControl
200 μm
200 μm
shGPR81
h
200 μm
D
Merge
E
C
shGPR81
hG 81
200 μm
F
ASPC1-low
200 μm
ASPC1-high
200 μm
Supplementary Figure 1: Lentiviral shRNA mediated knockdown or ectopic expression of
GPR81.
(A) Relative mRNA expression of GPR81 in Capan-II cells following lentiviral stable knockdown of
GPR81 (shGPR81) or lentiviral control (shControl). Results are the mean ± SEM, normalized to
shControl. n ≥ 3. ***p<0.001; ****p<0.0001 by ANOVA.
(B) shControl (top) or shGPR81 (bottom) Capan-II cells were coimmunostained for GPR81 (red) and
DAPI (blue). Scale bar, 200μm. n ≥ 3.
(C) Quantification of GPR81 protein levels, normalized to shControl. Data are mean ± SEM. n ≥ 3.
**p<0.01 by t-test.
(D) Relative mRNA expression GPR81 of BxPC3 cells transfected with siControl or siGPR81. Data are
p
by
y ANOVA.
mean ± SEM, normalized to siControl. n ≥ 3, ***p<0.001
(E) Relative mRNA expression of GPR81 mRNA in ASPC1-low and ASPC1- high cells. Data are
mean ± SEM, normalized to ASPC1-low cells. **p<0.01 by t-test.
(F) ASPC1-low (top) or ASPC1-high (bottom) cells were immunostained for GPR81 (red). Scale bar,
200μm. n ≥ 3.
Supplementary Figure 2
Supplementary Figure 2: Lentiviral shRNA mediated knockdown of GPR81 is associated with
reduced levels of MCT1. Relative mRNA expression of MCT1 in Capan-II cells following lentiviral
stable knockdown of GPR81 with 2 unique shRNA (shGPR81-1 & shGRP81-2) or lentiviral control
(shControl). Results are the mean ± SEM, normalized to shControl. n ≥ 3. ** p<0.01; ***p<0.001 by
ANOVA
Gene
Forward Primer
Reverse Primer
GPR81
AATTTGGCCGTGGCTGATTTC
ACCGTAAGGAACACGATGCTC
PGC1α
GCGGACAGAACTGAGAGACC
CCATCATCCCGCAGATTTAC
MCT1
CACTTAAAAATGCCACCAGCA
AGAGAAGCCGATGGAAATGA
MCT4
GTTGGGTTTGGCACTCAACT
GAAGACAGGGCTACCTGCTG
CD147
CCATGCTGGTCTCGAAGTCAG
CCGTTCATGAGGGCCTTGTC
18S
GAGCGGTCGGCGTCCCCCAACTTC
GCGCGTGCAGCCCCGGACATCTAA
Supplemental Table 1: Primer sequences for quantitative RT-PCR