Supplementary Figure Legends

Supplementary figure 1: Epithelial and stromal marker expression in human prostate-derived patient-matched cells, BPH-1 and WPMY-1 cells: E- cadherin and Vimentin expression in WFU primary cells. E/B= Epithelial/ benign, E/Ca= Epithelial/cancer. BPH-1 and WPMY-1 are epithelial and stroma-derived cell lines respectively.

Supplementary figure 2: Human Prostate-derived patient-matched cell lines exposed to metabolic inhibitors: MTS proliferation assay of patient-matched prostate-derived benign (A) and cancer (B) cells exposed to Etomoxir and/or Orlistat for 48 hours. Graphs represent Mean ± SD. b p<0.001 different from Etomoxir treatment in benign cells. c p<0.01, different form Orlistat+Etomoxir in benign cells.

Supplementary figure 3: Cleaved Caspase 3 expression in LNCaP and BPH-1 cells: Cells were treated with etomoxir over a 16 hour period and the activation of caspase 3 was examined by western blot in the cell lysates.

Supplementary figure 4: Palmitate supplementation decreases the spliced/unspliced XBP-1 ratio: LNCaP cells were treated over 16 hours with etomoxir and and with or without 100uM palmitate conjugated to albumin. Trizol was added to cells to isolate total RNA. cDNA was generated and used for qRTPCR to measure the spliced (sXBP1) and unspliced (unsXBP1) transcripts. The Y axis indicates the ratio of the normalized XBP1 gene expression. *p<0.05 compared to all other treatments. # p<0.05 compared to orl+eto without palmitate supplementation.

Supplementary Table 1: Primers used to analyze expression of the ER Stress-related genes and AR-related gene expression.

Supplementary Table 2: Antibodies used for immunoblots and IHC.

Supplementary Table 1: Primers
name / sequence / length
AR (all forms)-F / GAAAGCGACTTCACCGCAC / 20
AR (all forms)-R / AAAACATGGTCCCTGGCAGT / 21
ARfl-F / ACATCAAGGAACTCGATCGTATCATTGC / 28
ARfl-R / TTGGGCACTTGCACAGAGAT / 20
ARv7-F / CCATCTTGTCGTCTTCGGAAATGTTATGAAGC / 32
ARv7-R / TTTGAATGAGGCAAGTCAGCCTTTCT / 26
PSA-F / CGGATGCTGTGAAGGTCATGGAC / 23
PSA-R / GGGTCAAGAACTCCTCTGGTTC / 21
NKX3.1-F / GAGACGCTGGCAGAGACC / 18
NKX3.1-R / CTGAGTGTGGGAGAAGGCAG / 20
GADD34-F / GAGCCCTGCCCCTTCCGAGT / 20
GADD34-R / TCGGAGAAGCGCACCTTTCTGG / 22
ATF-4-F / TCCCCCTTCGACCAGTCGGG / 20
ATF-4-R / AGGCATCCTCCTTGCTGTTGTTGG / 24
CHOP-F / GCGACTCGCCGAGCTCTGATT / 21
CHOP-R / GGCCCAAGTGGGGGACTGATG / 21
HSPA5-F / GTGCAGAAACTCCGGCGCGA / 20
HSPA5-R / GACCGGAACAGATCCATGTTGAGC / 24
XBP-1-F / AAACAGAGTAGCAGCTCAGACTGC / 24
XBP-1-R / TCCTTCTGGGTAGACCTCTGGGAG / 24
B2M-F / AGGCTATCCAGCGTACTCCAAAGA / 24
B2M-R / TCGGATGGATGAAACCCAGACACA / 24
XBP-1u-F / GGTCTGCTGAGTCCGCAGCACT / 22
XBP-1s-F / GGTCTGCTGAGTCCGCAGCAGG / 22
XBP-1-R / GGGCTTGGTATATATGTGG / 19
RPL13A-F / CCTGGAGGAGAAGAGGAAAGAGA / 23
RPL13A_R / TTGAGGACCTCTGTGTATTTGTCAA / 25
Supplementary Table 2: Antibodies
Antibody / Catalogue number / Company
pEIF2a / 9791 / Cell Signaling
pAKT / 4051 / Cell Signaling
AKT / 9272 / Cell Signaling
pBAD (S136) / 5286 / Cell Signaling
pBAD (S112) / 9291 / Cell Signaling
pBAD (S155) / 9297 / Cell Signaling
c-caspase 3 / 9661 / Cell Signaling
mTOR signaling Kit
(mTOR, S6K 4EBP1) / 9862 / Cell Signaling
AMPK signaling kit
(AMPK, ACC) / 9957 / Cell Signaling
CPT-1A / 15184-1-AP / Proteintech
LC3 / NB100-2331 / Novus Biologicals
B-actin / 8226 / Abcam
HSP90 / 13495 / Abcam
Goat-anti Mouse-HRP / SC-2064 / Santa Cruz
Goat-anti Rabbit-HRP / SC-2030 / Santa Cruz
p-Ribosomal-S6 / 4858 / Cell Signaling