Supporting Information
Supplemental Material, Table S1
Table S1. Primers used for quantitative real-time polymerase chain reaction (PCR)
Gene
Primers
Sequences(5’→3’)
IL-1β
Forward
CACGGAGTTTCTTCTGGCCT
Reverse
CAATTCGGATCTGCAGCACG
Forward
GCCAATGGGGCACTGAA
Reverse
TGGTGGTGAAGAAGAGGAACAA
Forward
CACAAACCGGAGGCCTTTTG
Reverse
CATGCTGTAGGCCCCAAAGA
Forward
AGAGTCTGCTACAGCCCTGA
Reverse
CAGATTTGGGAAGGCGTCCA
Forward
TTGAAGACGTACCCTGTGCC
Reverse
GAGGCCATGAGCTCCTTGTT
Forward
GTCTAAATCGGGAGTTGGAATCA
Reverse
CACAGTATGACACAACAGCCCA
Forward
CACAAGAAAAATCCTTGGGTCAA
Reverse
GACAGCAGATTGCGACACACA
Forward
CCTAAGATAGCGGTTGATGG
Reverse
GACTTGAGAATGGATGCGAA
Forward
CGCTGGCGTCCGCCTGATCCTCA
Reverse
ACGCACAGCCGCCAGTAGGTAGAT
Forward
TGAACAGCTTCCGGATGATG
Reverse
TGAAGCCTCTCTTCTCCTCGAT
Forward
GTCAGATGAAAGCAAAGCGA
Reverse
CATAGTTGGGCTCAGGGTCT
Forward
ATCAGCCTCTGGAATGAAGGGG
Reverse
GCAGCCAATGTTGAAGCAAATCC
Forward
GCACCGTCAAGGCTGAGAAC
Reverse
TGGTGAAGACGCCAGTGGA
IL-6
IL-10
TNFβ1
iNOS
COX-2
NOX1
NOX2
p22phox
p40phox
p47phox
p67phox
GAPDH
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Figure S1. The effect of PM2.5 on cell viability of K562 cells. (a) MTT assay was used to
determine the cell viability of K562 cells with PM2.5 exposure. (b) Morphological photo of cells
were shown. Cells were examined with light microscope (10×). Values were representative of at
least three biologically independent experiments with similar results. *p < 0.05, **p < 0.01,
compared to controls.
2
Figure S2. The effect of PM2.5 on the localization of NF-κB in nucleus. The localization of
NF-κB p65 through indirect immunefluorescence using FITC conjugated secondary antibody. The
nucleus was stained with DAPI. Cells viewed at 60×magnification (Delta Vasion, USA).
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Figure S3. PM2.5 activates the NADPH oxidase. RT-PCR assay was carried out to measure the
expressions of NAPDH oxidase subunits. (a) The effects of PM2.5 on NOX1, NOX2 and p22phox
expressions in HL-60 cells. (b) The effects of PM2.5 on p40phox, p47phox and p67phox expressions in
HL-60 cells. (c) The mRNA levels of NOX1, NOX2 and p22phox expressions in K562 cells upon
PM2.5 exposure. (d) The mRNA levels of p40phox, p47phox and p67phox expressions in K562 cells
upon PM2.5 exposure. Data represented were mean ± SD of three identical experiments made in
four replicates. *Statistically significant differences as compared to controls (*p < 0.05, **p <
0.01).
4
Figure S4. The effect of NAC on PM2.5-enhanced localization of NF-κB in nucleus. After
co-treatment with NAC, the localization of NF-κB p65 through indirect immunofluorescence
using FITC conjugated secondary antibody. The nucleus was stained with DAPI. Images were
captured by a fluorescence microscope and viewed at 60×magnification (Delta Vasion, USA).
5