1. Prognostic Significance of Fbw7 in Human Melanoma and Its Role in Cell Migration 
Yabin Cheng, Guangdi Chen, Magdalena Martinka, Vincent Ho, Gang Li 
Journal of Investigative Dermatology 
Volume 133, Issue 7, Pages (July 2013)
DOI: /jid
Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

2. Figure 1
Fbw7 expression is reduced in human advanced melanoma. (a–d) Representative images of Fbw7 immunohistochemical staining in human melanoma TMA. (a, c) Normal nevus with positive staining. Arrows indicate positive Fbw7 nuclear staining. (b, d) Metastatic melanoma with negative staining. (a, b) Bar=40μm; (c, d) bar=10μm. (e) Reduced Fbw7 expression correlates with melanoma progression. DN, dysplastic nevi; Fbw7, F-box and WD repeat domain-containing 7; MM, metastatic melanoma; NN, normal nevi; PM, primary melanoma; TMA, tissue microarray. *P<0.05, **P<0.01.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

3. Figure 2
Kaplan–Meier survival analyses of melanoma patients. Patients with negative Fbw7 (F-box and WD repeat domain-containing 7) expression have a significantly worse (a) overall and (b) disease-specific 5-year survival than those with positive Fbw7 expression (P=0.037 and 0.015, respectively, log-rank test).
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

4. Figure 3
Fbw7 (F-box and WD repeat domain-containing 7) mRNA expression in normal melanocytes and melanoma cell lines. (a) Fbw7α protein expression level was reduced in melanoma cell lines as compared with normal melanocytes. (b) Total Fbw7 mRNA level was reduced in melanoma cell lines as compared with melanocytes. (c) Comparison of mRNA expression of three Fbw7 isoforms in melanocytes and melanoma cell lines.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

5. Figure 4
Enforced Fbw7α (F-box and WD repeat domain-containing 7) expression inhibits melanoma cell migration. (a) Representative images of the effects of Fbw7α overexpression on melanoma cell migration. (b) The inhibition of Fbw7α on melanoma cell migration was quantified by counting the migrated cells in five random fields of each well; error bars equal to means±SD. The data were obtained from three independent experiments. **P<0.01. (c) Western blot analysis of Fbw7α expression, total extracellular signal–regulated kinase 1/2 (ERK1/2), and phosphorylated ERK1/2 (p-ERK1/2).
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

6. Figure 5
Treatment with selective MEK (MAPK/ERK kinase) inhibitor, PD98059, abolished Fbw7α (F-box and WD repeat domain-containing 7) knockdown–induced melanoma cell migration. (a) Representative images show the effects of PD98059 on melanoma cell migration. After Fbw7α knockdown, MMRU cells were treated with 80μM PD98059 for 24hours in serum-free medium, and then the wound-healing assay was performed. (b) Quantification of a. The induction of Fbw7α knockdown on melanoma cell migration was quantified by counting the migrated cells in five random fields of each well; error bars equal to means±SD. Data were obtained from three independent experiments. ***P< (c) Western blot analysis showed efficient Fbw7α knockdown using two small interfering RNAs (siRNAs). (d) Western blot analysis of extracellular signal–regulated kinase 1/2 (ERK1/2) phosphorylation (p-ERK1/2) by MEK inhibitor PD98059.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

7. Figure 6
Fbw7α (F-box and WD repeat domain-containing 7) affects RhoA activity and stress fiber formation. (a) Fbw7α-knockdown (KD)–induced RhoA activity was abolished by PD MMRU cells transfected with siControl and siFbw7α small interfering RNAs (siRNAs) were serum starved for 24hours, and had serum stimulation for 30minutes. (b) Decreased stress fiber formation in Fbw7α-overexpressed MMRU cells. MMRU cells were transfected with FLAG-Fbw7α or vector control, followed by serum starvation for 24hours and serum stimulation for 30minutes. DAPI, 4',6-diamidino-2-phenylindole. (c) Increased stress fiber formation after Fbw7α-KD. MMRU cells were transfected with siControl and siFbw7α siRNAs, followed by serum starvation for 24hours and serum stimulation for 30minutes. For MEK (MAPK/ERK kinase) inhibitor treatment, MMRU cells were incubated with serum-free medium containing 80μM PD98059 for 24hours. Original magnification × 630. (d) Quantification of b and c. Relative intensity of F-actin was examined by ImageJ software; **P<0.01, ***P<0.001.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions