Table S1. Primers used for quantitative PCR amplification of genes for plasma and matrix proteins
and lubricant in synoviocytes from rabbit (Oryctolagus cuniculus). All primers were designed as
unique among all mRNA reference sequences in rabbit by Blast search, but may be shared among
alternatively spliced variants.
Genes
GenBank ID #
Forward sequence (5’ to 3’)
Reverse sequence (5’ to 3’)
qPCR products *
CT
MP
Size (bp)
A2M
XM_008259802 GCGTGGTTTGGCAGACATTT
ATGAAGCTCCTCAAGCCACC
>36
GCACCTCTTGCTGTTGCTTC
AAGAGAGACGATGGGGGACA
34
GGAGGCTTCTCCTCTACCCA
GATGTGGGTCGTTTCCCAGT
36
73
395
C3
NM_001082286 AGGCCACCTTCATGGGTTTC
GCTTCTTCACTGTCTCAGGGG
>36
CGGAGAAAGAGGACGGGATG
TGTTGGGTTTCTCGCCCAC
33
84
369
A1AT NM_001171081 CACAAGGTGGTGCTGACCAT
CTTTAATGTGCTGCGGTGGG
28
80
209**
AHSG XM_002716457.2 CGTGTTGTCTGCGAAGTGTG
ACAGGTCACCTCCACCTCTT
33
73
384
GCTGGCGGACAAGCAATATG
ACAGGTCACCTCCACCTCTT
>36
TGCCCAGGGAAGGTCAGATA
GTGAGGTCGATTGGAAGGCA
33
271**
SPP2 XM_008275266 TTCCCCGTGTACGACTACGA
TTCTCAGCGAGCTTCCGAAC
28
80
209**
IGF1
NM_001082026 CATCCTGTCCTCCTCGCATC
CCGTATCCTGTGGGCTTGTT
31
76
165
GTGTGGAGACAGGGGCTTTT
TCATCCACGATGCCTGTCTG
>36
AACAAGCCCACAGGATACGG
TCCAGCCTCCTCAGATCACA
31
82
98**
TGTGATCTGAGGAGGCTGGA
TCTGAGTCTTGGGCATGTCG
31
84
103
CP
XM_008266309 CGGTACATTTTCACGGCCAC
TGCTCGCTGTGAATGGATAGTT 33
80
255
TF
NM_001101694 TTATGAGTACGTCACCGCCG
CCTTCCTCCGGTGCTTTCTT
>36
CAGAACACTGGTGGACGGAA
GTGTTTTTGCCCCGAAGGTC
>36
81
C1INH XM_008271443 CACCCCCTGGAGGACATAGA
GAAGATCCGGGTCATCCGTC
31
79
CRP
NM_001082265 CCCTGTGGATCCAACTCACC
TCCCACCAGCATAGACGGTA
26
80
269**
PRG4 NM_005807.3
ACACGTCTACCACCCAACAC
TGGAAGACTGGGTCTGGGAT
36
75
AACTACAACTGCTGCACCTAAGA TTGATGTCATCTTGCGGGGA
>36
MMP1 NM_002421.3
CCACAAACCCCAAAAGCGTG
TGTCCTTGGGGTATCCGTGT
>36
CATGGTGTGAGTCCAAAGAAGG GCTCTCTGGGATCAACGTCA
>36
TIMP1 NM_003254.2
TCTGCAATTCCGACCTCGTC
TGCATTCCTCACAGCCAACA
>36
TCTGCAATTCCGACCTCGTC
TGCATTCCTCACAGCCAACA
>36
HAS1 XM_008249800 CCTTAACCCCCTGGACTCCT
CCGCTGATGCAGGATACACA
25
81
114**
HAS2 NM_001082010 CTGGGACGAAGCGTGGATTA
ATGCACTGGACACACCCAAA
21
77
239**
HAS3 NM_001082709 ATCCCCAAGTAGGGGGAGTC
CAGCCAAAGTAGGACTGGCA
>36
GCCAGTCCTACTTTGGCTGT
ACCGACTCGTAGGTCATCCA
>36
CCATCTGGGTGGCAGTTCTT
CCTCAGCAAAGGCCAAGCTA
30
80
204**
SAA4 NM_199161.3
ACAGCACAGATCAGCACCAT
CAAAAGCCTCGCCAAGGAAC
>36
GTTCCTTGGCGAGGCTTTTG
CTGATCACTTCTGCAGCCCA
>36
GATTCTTTGGCCATGGTGCG
ATTGGGGTCTTTGCCACTCC
>36
GGAGTGGCAAAGACCCCAAT
TCTCCCCGCTTTGTATCCCT
34
80
127
18S
NR_003286
CGGGTCATAAGCTTGCGTT
CCGCAGGTTCACCTACGG
17
84
204**
*All values were obtained by amplification of cDNAs reverse transcribed with random primers from RNA of untreated
synoviocytes. CT, cycle count; MP, melting point of amplicon. Cutoff values for meaningful CTs were set at 33 for room of
possible decreases (higher and unreliable CT values) under certain treatments. **Primer sets were used in the calculation and
presentation of Figure 5.
Figure S1. Purification and sequencing of high molecular band from cultured medium from Hig82
cells. Serum-free medium after culturing Hig-82 cells for 16 hrs (25 ml pooled from 5 plates of 10 cm
diameters) was concentrated on Centricon-10 to ~100 l. The sample was mixed with non-reducing LB
and loaded in 4 lanes and separated on a 4-20% SDS-PAGE. One lane was stained with Coomassie blue,
and a band corresponding to 360-720 kDa three unstained lanes was excised, inserted into a new
Centricon-10 concentrator, and electro-eluted in transfer buffer in a Hoeffer apparatus. The sample was
concentrated again to ~50 l, mixed with 25 l 3x reducing LB, and boiled for 15 min. The sample was
separated again in 4-20% SDS-PAGE and stained. Gel slices were cut, digested with trypsin, and
analyzed for proteins with LC-MS/MS.
List of Abbreviations:
A1AT1-antitrypsin or serpinA1
A2M, 2-macroglobulin
ACE2, angiotensin I converting enzyme 2
C1-antichymotrypsin or serpinA3
AEBSF, 4-(2-Aminoethyl)benzenesulfonyl fluoride hydrochloride
AFP, -fetoprotein
AHSG, 2-HS-glycoprotein or fetuin A
ALB, albumin
ALLN, N-[N-(N-acetyl-L-leucyl)-L-leucyl]-L-norleucine
AT3, antithrombin III or serpinC1
MSH, -mercaptoethanol
BFA, brefeldin A
BMP-2, bone morphogenetic protein-2
BRINP3, Bone morphogenetic protein/retinoic acid inducible neural-specific 3
C1INH, C1 inhibitor or serpinG1
C1S, Complement component 1, s subcomponent
C8A, complement component 8,  polypeptide
C3, complement C3
CGN, cis-Golgi network
CP, ceruloplasmin
COL, collagen
CRP, C-reactive protein
CSF1, macrophage colony-stimulating factor 1
CTRP7, C1q and tumor necrosis factor related protein 7
DCI, 4-dichloroisocoumarin
DCN, decorin
DTT, dithiothreitol
dMJ, deoxymannojirimycin
dNJ, 1-deoxynojirimycin
E64d, 2S,3S-trans-(ethoxycarbonyloxirane-2-carbonyl)-L-leucine-(3-methylbutyl) amide
EDTA, Ethylenediaminetetraacetic acid
EGTA, ethylene glycol tetraacetic acid
ER, endoplasmic reticulum
FCS, fetal calf serum
FMOD, fibromodulin
FN, fibrinogen
HAS 1, 2, or 3, hyaluronan synthase-1, 2, or 3
Hg, haptoglobin
Hig-82, rabbit (Oryctolagus cuniculus) synoviocytes
HSPG2, heparan sulfate proteoglycan-2
IAM, iodoacetamide
IFNG, interferon, 
IGF-1, insulin-like growth factor-1
IGSF8, immunoglobulin superfamily member 8
IIGBP1, Interferon-induced guanylate-binding protein 1
IL1, interleukin 1
IL6, interleukin 6
IL33, interleukin 33
ITGA11, integrin, 11
ITIH4, inter--trypsin inhibitor H4
LAMC1, Laminin, 1 (formerly LAMB2)
LB, loading buffer
MG-132, carbobenzoxy-L-leucyl-L-leucyl-L-leucinal
MMP, matrix metalloproteinase
NEM, N-ethylmaleimide
OA, osteoarthritis
PAMR1, Peptidase domain containing associated with muscle regeneration 1
PICA, porcine inhibitor of carbonic anhydrase
PRG4, proteoglycan 4 or lubricin
PVDF, polyvinylidene difluoride
PZP, pregnancy zone protein
qPCR, quantitative polymerase chain reaction
RA, rheumatoid arthritis
RPF2, rabbit permeability factor-2
SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis
SF, synovial fluid
Serpin, serine protease inhibitor
SPINK5, Serine peptidase inhibitor, Kazal type 5
Spp24, secreted phosphoprotein 24 kD
SVEP1, Sushi, von Willebrand factor type A, EGF and pentraxin domain containing 1
SW, swainsonine
TF, transferrin
TIMP, metalloproteinase inhibitor
TGF-1, transforming growth factor-beta 1
TM, tunicamycin
TNFtumor necrosis factor alpha
TTR, transthyretin