Supplementary Figure 1. Determination of minimal erythema dose (MED) in UVB irradiated hairless mice. The minimal skin erythema was observed on skin area irradiated with 120 mJ/cm2 (1.2 mW/cm2 x 100 s) UVB.

Supplementary Figure 2. The anti-wrinkle effects of DIC, an elastase inhibitor. DIC did not attenuate wrinkle formation and degradation of collagen fibers by UVB (a) Replica photography (b) Massons’s trichrome staining for collagen fibers (c) Verhoeff staining for elastin fibers. Magnification: x 200. Bar: 200 ㎛ .

(a)UVB (13 W)

Non-UV Vehicle DIC (0.01%)

(b)

(c)

Supplementary Figure 3. The inhibitory effects of KPA, chymostatin and DIC on fibronectin fragmentation and conversion of proMMP-1 to active MMP-1 in cathepsin G (CG) treated NHFs. The fragmentation of Fn was analyzed in presence of KPA, chymostatin and DIC by western blot using anti-fibronectin Ab. (a, c) Specific inhibitors of cathepsin G like KPA and chymostatin prevented Fn degradation and conversion of proMMP-1 to active MMP-1 by cathepsin G (18). (b, d) DIC, an elastase specific inhibitor, did not prevented Fn fragmentation and conversion of proMMP-1 to active MMP-1 by cathepsin G.

191

97

64

51

39

2819

(kDa)LC

KPA(µM) Chymostatin (µg/ml) DIC (µM) - 0.8 0.16 - 10 4 - 0.8 0.16- + + + + + + - + + +

Intact Fn

Fn-fr’s

CG (1mU/ml)

CG (1mU/ml)

Chymostatin KPA DIC (µg/ml) (µM) (µM) 10 4 2 0.8 0.16 0.0032 0.8 0.16 - - + + + + + + + + + + +

Pro MMP-1

Active MMP-1

(a) (b)

(c) (d)

Pro MMP-1

Active MMP-1

Con CG (mU) - 1 5 10

Con Elastase (mU) - 1 5

Supplementary Figure 4. Conversion of proMMP-1 to active MMP-1 by cathepsin G (CG) or elastase treatment in NHFs. Cathepsin G promoted the conversion of proMMP-1 to active MMP-1, but elastase did not.

Supplementary Figure 5. Pictures of 3D dermal equivalent (3D DE). 3D DE contains collagen, fibronectin and normal human fibroblasts in DMEM medium. It changes from liquid state to gel state after incubation at 37 for 2 hour. (a) 3D DE in ℃24 well plates and (b) gel state 3D DE out 24 well plates.

(a) (b)

Supplementary Figure 6. Verhoeff staining of elastic fibers after KPA treatment. (a) Non UV control (b) UVB-irradiated vehicle treated group (c) UVB-irradiated retinoic acid treated group (d) UVB-irradiated KPA treated group. Magnification: x 200. Bar: 200 ㎛

(a) (b)

(c) (d)

Non UV UVB+Vehicle

UVB+ RA (0.01%) UVB+ KPA (0.025%)