Journal: Diagnostics

Article Title: Alpha-Enolase (ENO1) Correlates with Invasiveness of Cutaneous Melanoma—An In Vitro and a Clinical Study

doi: 10.3390/diagnostics12020254

Figure Lengend Snippet: The expression level of ENO1 in the cell lysates from primary melanocytes and melanoma cell lines. Representative Western blots showing ENO1 and Akt 1/2/3 expression (for normalization) in protein lysates obtained from the primary human melanocytes (HEM) and indicated melanoma cell lines ( a ). Densitometric ENO1/Akt ratios are shown as mean values ( n = 3 except for HEM, n = 2) ± standard error of the mean (SEM) ( b ). The significance level was set at p = 0.001–0.0001 (***).

Article Snippet: Human epidermal melanocytes, adult (HEMa, 104−05A) and primary human epidermal melanocytes (lightly pigmented) (HEMn-LP, C0025C) were purchased from Cell Applications Inc (San Diego, CA, USA), and Cascade Biologics/Gibco (Carlsbad, CA, USA), respectively.

Techniques: Expressing, Western Blot

Journal: Oncogene

Article Title: RSK regulates activated BRAF signalling to mTORC1 and promotes melanoma growth

doi: 10.1038/onc.2012.312

Figure Lengend Snippet: Inhibition of ERK/RSK signalling decreases constitutive activation of mTORC1 in melanoma. (A) Four melanoma cell lines were used in this study. While A375 and Colo829 cells harbour a B-Raf V600E mutation, WM852 and WM1361 cells carry an N-Ras mutation at Q61 (R or K). (B) Phosphorylation of endogenous RSK, ERK1/2, rpS6, S6K1 and 4E-BP1 was monitored in total extracts from serum-starved melanoma cell lines and normal human melanocytes treated or not with insulin (100 nM) for 30 min. Cell lysates were also immunoblotted for total protein levels (RSK1, ERK1, rpS6, S6K1, 4E-BP1 and β-actin). (C) Serum-starved melanoma cells were treated with the indicated inhibitors for 60 minutes. Immunoprecipitated S6K1 kinase activity was assayed as in Fig. 1. (D) Phosphorylation of endogenous rpS6 and S6K1, and total rpS6 protein level were monitored by immunoblotting.

Article Snippet: The human epidermal melanocytic (105-25N) cell line was purchased from Cell Applications (San Diego, CA) and grown in Melanocyte Growth Medium (135-500, Cell Applications).

Techniques: Inhibition, Activation Assay, Mutagenesis, Phospho-proteomics, Immunoprecipitation, Activity Assay, Western Blot

Journal: Advances in Dermatology and Allergology/Postȩpy Dermatologii i Alergologii

Article Title: Effects of tea polyphenols on UVA-induced melanogenesis via inhibition of α-MSH-MC1R signalling pathway

doi: 10.5114/ada.2022.115890

Figure Lengend Snippet: Effects of TPS on the expression of α-MSH. Representative chart trace of α-MSH inhibition by 5, 10, and 15 μg/ml of the TPS on HaCaT cells ( A ) and HEM cells ( B ) with UVA exposure (15 J/cm 2 ) or not by ELISA. Error bars show means ± SEMs. * P < 0.05, ** p < 0.01, *** p < 0.001 versus non-treated cells, # p < 0.05, ## p < 0.01, ### p < 0.001 versus UVA-treated cells

Article Snippet: The following material was purchased from the following manufacturers: tea polyphenols (98% purity), Chinese Academy of Agricultural Sciences, Tea Research Institute; 10% foetal bovine serum (FBS), DMEM and DMSO, Gibco/BRL (Grand Island, NY, USA); Bovine serum albumin (BSA), levodopa (L-DOPA) and nonapeptide-1 acetate salt (N-1A), MedChemExpress (Shanghai, China); and α-MSH ELISA Kit, CUSABIO (Wuhan, China).

Techniques: Expressing, Inhibition, Enzyme-linked Immunosorbent Assay

Journal: Advances in Dermatology and Allergology/Postȩpy Dermatologii i Alergologii

Article Title: Effects of tea polyphenols on UVA-induced melanogenesis via inhibition of α-MSH-MC1R signalling pathway

doi: 10.5114/ada.2022.115890

Figure Lengend Snippet: TPS inhibited melanogenesis through suppressing the α-MSH-MC1R signalling pathway in HaCaT cells and HEM cells. A, B – HEM was pretreated or not with UVA exposure (15 J/cm 2 ) before TPS (10 μg/ml) and N-1A (20 μm) were applied. Melanin contents and the expression of tyrosinase were measured as described in methods. C – HaCaT cells were treated with TPS (10 μg/ml) and/or N-1A (20 μm) in the presence or absence of UVA exposure (15 J/cm 2 ). ELISA was then applied to detect the expression of α-MSH. D – HEM cells were treated with TPS (10 μg/ml) and N-1A (20 μm) in the presence or absence of UVA exposure (15 J/cm 2 ). ELISA was then applied to detect the expression of α-MSH. * P < 0.05, ** p < 0.01, *** p < 0.001 versus non-treated cells, # p < 0.05, ## p < 0.01, ### p < 0.001 versus UVA-treated cells, a p < 0.05, aa p < 0.01, aaa p < 0.001, ns p > 0.05

Article Snippet: The following material was purchased from the following manufacturers: tea polyphenols (98% purity), Chinese Academy of Agricultural Sciences, Tea Research Institute; 10% foetal bovine serum (FBS), DMEM and DMSO, Gibco/BRL (Grand Island, NY, USA); Bovine serum albumin (BSA), levodopa (L-DOPA) and nonapeptide-1 acetate salt (N-1A), MedChemExpress (Shanghai, China); and α-MSH ELISA Kit, CUSABIO (Wuhan, China).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay