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Figure Legends

Fig. 1. Microarray analysis of gene expression profiling

Microarray profiling of chondrocytes showed genes that are commonly or conflictingly

regulated by CTS and µG (n = 1 per group). Category I: genes upregulated by both CTS and

µG, Category Ⅱ: genes upregulated by CTS and downregulated by µG, Category Ⅲ: genes

upregulated by µG and downregulated by CTS, and Category Ⅵ: genes downregulated by

both CTS and µG.

Fig. 2. Fat1 mRNA gene expression in response to CTS and µG

The graphs show the expression of Fat1 in each group. Data are shown as the mean ± SD of 4

wells per group. *P < 0.05.

Fig. 3. Immunocytochemical staining of β-tubulin

Representative images of immunocytochemical staining shows (A-D) β-tubulin.in each group

Scale bars = 50 µm.

Fig. 4. Immunocytochemical staining of F-actin and Fat1

Representative images of immunocytochemical staining shows (A-B) F-actin, (E-H) Fat1, (IL) nucleus staining-DAPI, and (M-P) Merge in each group. Scale bars = 50 µm. The graph

26

right shows staining intensity of (Q and R) F-Actin in whole cell region (n =150-200 cells per

group), (S and T) Fat1 in whole cell region (n = 60-70 cells per group), and (U and V) Fat1 in

the cell cortical region (n =60-70 cells per group) in each group. Data are shown as the mean

± SD. *P < 0.05.

27

Figure 1

28

Figure 2

29

Figure 3

30

Figure 4

31

Table 1. GO terms of genes upregulated by both CTS and µG

symbol

description

GO biological process

Traf3ip3

TRAF3 interacting protein 3

GO:0008150 (biological process)

S100g

S100 calcium binding protein G

Unavailable annotation

Kell blood group precursor

(McLeod phenotype) homolog

GO:0006865 (amino acid transport)

GO:0006874 (cellular calcium ion homeostasis)

GO:0008361 (regulation of cell size)

GO:0010961 (cellular magnesium ion homeostasis)

GO:0042552 (myelination)

GO:0048741 (skeletal muscle fiber development)

prokineticin receptor 1

GO:0007165 (signal transduction)

GO:0007186 (G-protein coupled receptor protein signaling pathway)

GO:0007623 (circadian rhythm)

GO:0043066 (negative regulation of apoptotic process)

GO:0060976 (coronary vasculature development)

Xk

Prokr1

32

Table 2. GO terms of genes upregulated by CTS and downregulated by µG

symbol

description

GO biological process

GO:0002088 (lens development in camera-type eye)

GO:0003382 (epithelial cell morphogenesis)

GO:0003412 (establishment of epithelial cell apical/basal polarity

involved in camera-type eye morphogenesis)

GO: 0007015 （actin filament organization）

GO:0007155 (cell adhesion)

Fat1

Il17rb

FAT tumor suppressor homolog 1

interleukin 17 receptor B

GO: 0007156（homophilic cell adhesion via plasma membrane adhesion

molecules）

GO: 0007163 (establishment and/or maintenance of cell polarity)

GO: 0016337 (cell-cell adhesion)

GO:0043010 (camera-type eye development)

GO:0045197 (establishment or maintenance of epithelial cell apical/basal

polarity)

GO:0048593 (camera-type eye morphogenesis)

GO:0050729 (positive regulation of inflammatory response)

GO:0098609 (cell-cell adhesion)

GO:0019221 (cytokine-mediated signaling pathway cytokine-mediated

signaling pathway)

GO:0032736 (positive regulation of interleukin-13 production)

GO:0032754 (positive regulation of interleukin-5 production)

GO:0050729 (positive regulation of inflammatory response)

33

Table 3. GO terms of genes upregulated by µG and downregulated by CTS

symbol

Sfrp2

description

GO biological process

secreted frizzled-related protein 2

GO:0001569 (branching involved in blood vessel morphogenesis)

GO:0001756 (somitogenesis)

GO:0002063 (chondrocyte development)

GO:0003214 (cardiac left ventricle morphogenesis)

GO:0007584 (response to nutrient)

GO:0010975 (regulation of neuron projection development)

GO:0010719 (negative regulation of epithelial to mesenchymal transition)

GO:0010667 (negative regulation of cardiac muscle cell apoptotic process)

GO:0010950 (positive regulation of endopeptidase activity)

GO:0021915 (neural tube development)

GO:0030111 (regulation of Wnt signaling pathway)

GO:0030178 (negative regulation of Wnt signaling pathway)

GO:0030199 (collagen fibril organization)

GO:0030514 (negative regulation of BMP signaling pathway)

GO:0003151 (outflow tract morphogenesis)

GO:0031668 (cellular response to extracellular stimulus)

GO:0035567 (non-canonical Wnt signaling pathway)

GO:0036342 (post-anal tail morphogenesis)

GO:0042493 (response to drug)

GO:0042733 (embryonic digit morphogenesis)

34

GO:0045600 (positive regulation of fat cell differentiation)

GO:0042662 (negative regulation of mesodermal cell fate specification)

GO:0043508 (negative regulation of JUN kinase activity)

GO:0046546 (development of primary male sexual characteristics)

GO:0048546 (digestive tract morphogenesis)

GO:0050732 (negative regulation of peptidyl-tyrosine phosphorylation)

GO:0060028 (convergent extension involved in axis elongation)

GO:0060349 (bone morphogenesis)

GO:0061056 (sclerotome development)

GO:0061185 (negative regulation of dermatome development)

GO:0071425 (hematopoietic stem cell proliferation)

GO:0071481 (cellular response to X-ray)

GO:0090175 (regulation of establishment of planar polarity)

GO:0090179 (planar cell polarity pathway involved in neural tube closure)

GO:0090244 (Wnt signaling pathway involved in somitogenesis)

GO:1902042 (negative regulation of extrinsic apoptotic signaling pathway

via death domain receptors)

GO:1904956 (regulation of midbrain dopaminergic neuron differentiation)

GO:2000035 (regulation of stem cell division)

GO:2000041 (negative regulation of planar cell polarity pathway involved

in axis elongation)

Gpr12

G-protein coupled receptor 12

GO:0006874 (calcium ion homeostasis)

GO:0007165 (signal transduction)

GO:0007186 (G-protein coupled receptor protein signaling pathway)

35

GO:0019222 (regulation of metabolic process)

Slc8a1

solute carrier family 8

(sodium/calcium exchanger),

member 1

GO:0001892 (embryonic placenta development)

GO:0002026 (regulation of the force of heart contraction)

GO:0002027 (regulation of heart rate)

GO:0002028 (regulation of sodium ion transport)

GO:0006874 (cellular calcium ion homeostasis)

GO:0006883 (cellular sodium ion homeostasis)

GO:0007154 (cell communication)

GO:0007584 (response to nutrient)

GO:0010763 (positive regulation of fibroblast migration)

GO:0010881 (regulation of cardiac muscle contraction by regulation of the

release of sequestered calcium ion)

GO:0014829 (vascular associated smooth muscle contraction)

GO:0021537 (telencephalon development)

GO:0030001 (metal ion transport)

GO:0030501 (positive regulation of bone mineralization)

GO:0033198 (response to ATP)

GO:0034614 (cellular response to reactive oxygen species)

GO:0035050 (embryonic heart tube development)

GO:0035902 (response to immobilization stress)

GO:0035994 (response to muscle stretch)

GO:0036376 (sodium ion export across plasma membrane)

GO:0042493 (response to drug)

GO:0042542 (response to hydrogen peroxide)

36

GO:0044557 (relaxation of smooth muscle)

GO:0048747 (muscle fiber development)

GO:0051481 (negative regulation of cytosolic calcium ion concentration)

GO:0051924 (regulation of calcium ion transport)

GO:0055013 (cardiac muscle cell development)

GO:0055074 (calcium ion homeostasis)

GO:0060048 (cardiac muscle contraction)

GO:0060402 (calcium ion transport into cytosol)

GO:0070509 (calcium ion import)

GO:0071313 (cellular response to caffeine)

GO:0071320 (cellular response to cAMP)

GO:0071901 (negative regulation of protein serine/threonine kinase

activity)

GO:0086036 (regulation of cardiac muscle cell membrane potential)

GO:0086064 (cell communication by electrical coupling involved in cardiac

conduction)

GO:0098703 (calcium ion import across plasma membrane)

GO:0098719 (sodium ion import across plasma membrane)

GO:0098735 (positive regulation of the force of heart contraction)

GO:0099566 (regulation of postsynaptic cytosolic calcium ion

concentration)

GO:1901660 (calcium ion export)

37

Fscn2

Dnahc6

Krt8

fascin homolog 2, actin-bundling

protein, retinal

GO:0007163 (establishment or maintenance of cell polarity)

GO:0042462 (eye photoreceptor cell development)

GO:0051017(actin filament bundle assembly)

dynein, axonemal, heavy chain 6

GO:0003341 (cilium movement)

GO:0007018(microtubule-based movement)

GO:0060285 (cilium-dependent cell motility)

keratin 8

GO:0000904 (cell morphogenesis involved in differentiation)

GO:0033209 (tumor necrosis factor-mediated signaling pathway)

GO:0045214 (sarcomere organization)

GO:0051599 (response to hydrostatic pressure)

GO:0051707 (response to other organism)

GO:0060706 (cell differentiation involved in embryonic placenta

development)

GO:0097191 (extrinsic apoptotic signaling pathway)

GO:0097284 (hepatocyte apoptotic process)

2210403B10Rik RIKEN cDNA 2210403B10 gene

GO:0007165 (signal transduction)

Olfr1218

GO:0007165 (signal transduction)

GO:0007186 (G-protein coupled receptor protein signaling pathway)

GO:0007608 (sensory perception of smell)

olfactory receptor 1218

GO:0050911 (detection of chemical stimulus involved in sensory

perception of smell)

Prap1

proline-rich acidic protein 1

Unavailable annotation

38

AK122525

Phospho1

cDNA sequence AK122525

Unavailable annotation

phosphatase, orphan 1

GO:0001958 (endochondral ossification)

GO:0030500 (regulation of bone mineralization)

GO:0035630 (bone mineralization involved in bone maturation)

39

Table 4. GO terms of genes downregulated by both CTS and µG

symbol

description

GO biological process

Serpina3k

serine (or cysteine) peptidase

inhibitor, clade A, member 3K

GO:0010466 (negative regulation of peptidase activity)

GO:0010951 (negative regulation of endopeptidase activity)

GO:0034097 (response to cytokine)

GO:0043434 (response to peptide hormone)

Klhl4

kelch-like 4 (Drosophila)

GO:0008150 (biological_process)

Me3

malic enzyme 3, NADP(+)dependent, mitochondrial

GO:0006090(pyruvate metabolic process)

GO:0006108(malate metabolic process)

1700093K21Rik RIKEN cDNA 1700093K21 gene

GO:0008150 (biological_process)

4930528A17Rik RIKEN cDNA 4930528A17 gene

Unavailable annotation

Gsta3

glutathione S-transferase, alpha 3

GO:0001657 (ureteric bud development)

GO:0006749 (glutathione metabolic process)

GO:0006805 (xenobiotic metabolic process)

GO:0046223 (aflatoxin catabolic process)

Slamf7

SLAM family member 7

GO:0002250 (adaptive immune response)

GO:0032814 (regulation of natural killer cell activation)

GO:0045087 (innate immune response)

Slitrk4

SLIT and NTRK-like family,

member 4

GO:0007409 (axonogenesis)

GO:0050807 (egulation of synapse organization)

GO:0051965 (positive regulation of synapse assembly)

GO:1905606 (regulation of presynapse assembly)

40

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