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Supplementary Information
Legends to supplementary figures
Supplementary Figure 1. Growth inhibitory effect and of MA1. Human PCa cell lines PC3 and
LNCaP were cultured at 96-well plates 24 h before treatment with 50 to 400 nM MA1 for 24
h. The viability was determined by the MTT assay. Values are means ± SD of three
experiments. *P < 0.05, versus MA1-untreated control group respectively.
Supplementary Figure 2. Apoptosis in PC3 and LNCaP cells as visualized by DAPI staining
after MA1 treatment. Cells were exposed for indicated times to 150 nM MA1 prior to
staining with DAPI for 30 min.
Supplementary Figure 3. Response of fluorescence probe coumarin derived azide 51. PC3
and LNCaP cells exposed to 150 nM MA1 or vehicle for 4 h were incubated with 30 μM probe
51 for 1 h in the dark and then examined by confocal microscopy (scale bar, 10 μm).
Supplementary Figure 4. Effect of MA1 on DNA damage and ER stress in PCa cells. a DNA
damage mediated by MA1 was estimated through the levels of γH2AX and p-BRCA1 by
western blotting assay response to MA1. b Induction of ER stress by MA1. The protein levels
of GRP78, CHOP, ATF4, p-PREK were tested by western blotting. c QRT-PCR analysis of
mRNA levels of GRP78, CHOP, ATF4, ATF3 and DDIT4. In a-c, PC3 and LNCaP cells were
treated with 150 nM MA1 of different times as indicated. GAPDH served as a loading
control.
Supplementary Figure 5. Effects of MA1 treatment on lysosomal stability. PC3 and LNCaP
cells were exposed to 150 nM MA1 for indicated times, and lysosomal stability was analyzed
by AO-uptake methods.
Supplementary Table 1.
Primers for quantitative RT-PCR analysis
Forward primer (5'->3')
Reverse primer (5'->3')
SOD2
TTCTGGACAAACCTCAGCCCTA
AACCTGAGCCTTGGACACCA
GSTP1
CCTGTACCAGTCCAATACCATCC
TCCTGCTGGTCCTTCCCATA
DJ-1
GTAGCCGTGATGTGGTCATTT
CTGTGCGCCCAGATTACCT
LC3B
GAAGATGTCCGACTTATTCGAGAG
ACTCTCATACACCTCTGAGATTGG
GRP78
GAAAGAAGGTTACCCATGCAGT
CAGGCCATAAGCAATAGCAGC
ATF3
TTGCAGAGCTAAGCAGTCGTGGTA
ATGGTTCTCTGCTGCTGGGATTCT
ATF4
TGGCCAAGCACTTCAAACCT
GTTGTTGGAGGGACTGACCAA
CHOP
GGAAACAGAGTGGTCATTCCC
CTGCTTGAGCCGTTCATTCTC
DDIT4
AGCGGCAGGACGCACTTGTC
GGCCGATCTGGGGTGGGAGT
GAPDH
TGGTCACCAGGGCTGCTT
AGCTTCCCGTTCTCAGCCTT