Fig 1: TCEAL7 increased the expression levels of AKT1, AKT2 and c-Myc in melanoma cells. a–d The mRNA and/or protein levels of AKT1, AKT2, p-AKT, c-Myc, STAT3, ß-catenin, TGF-ß, p-SMAD2, p-SMAD3, YAP and FOXO1 in Lenti-TCEAL7/si-TCEAL7-transfected WM-115 and A375 cells were determined by RT-PCR and western blotting assays. e–g Correlations between the expression levels of TCEAL7 and AKT1, AKT2 and c-Myc in melanoma samples were predicted by using the starBase database. h–j. Correlations between the expression levels of TCEAL7 and AKT1, AKT2 and c-Myc in 30 melanoma tissues samples were determined. (a and c *P < 0.05, compared with si-Scramble group; #P < 0.05, compared with Lenti-NC group; b and d, *P < 0.05, compared with Lenti-NC group; #P < 0.05, compared with si-Scramble group)
Fig 2: TCEAL7 inhibited the progression of melanoma through decreasing the expression of AKT1 and c-Myc. WM-115 and A375 cells were divided into Lenti-TCEAL7, Lenti-NC, Lenti-TCEAL7 + Lenti-AKT1/c-Myc and Lenti-AKT1/c-Myc groups and submitted to the following assays. a–d Cell proliferation was detected by using the CCK-8 assay. e, f Transwell chamber assay was used to detect cell invasion. g, h Flow cytometry assay was applied to detect cell apoptosis. i–l Scratch assay was used to assess cell migration ability. (*P < 0.05 and ***P < 0.001, compared with Lenti-NC group; #P < 0.05 and ###P < 0.001, compared with Lenti-TCEAL7 group)
Fig 3: miR-758-3p negatively modulated TCEAL7 expression. a BioVenn (https://www.biovenn.nl/index.php) was built to find the co-regulator of TCEAL7 from miRanda and miRDB diabetes. b RT-PCR analysis of the expression levels of miR-758-3p in melanoma tissues and nevus tissues (normal control). c RT-PCR was applied to determine miR-758-3p levels in normal melanocyte PIG1 and melanoma cell lines WM-115 and A375. d Kaplan–Meier was used to analyze the prognostic value of miR-758-3p in melanoma patients. e Correlation analysis between the expression levels of TCEAL7 and miR-758-3p in 30 melanoma tissues samples were determined. f RT-PCR analysis of the expression levels of miR-758-3p following cell infection with mimics-miR-758-3p or mimic-NC in WM-115 and A375 cells. g Western blotting analysis of the effect of miR-758-3p on the expression of TCEAL7 protein. h Putative binding sites between miR-758-3p and TCEAL7 3'UTR. i, j Dual-luciferase gene reporter system was used to assess the effect of miR-758-3p on the transcriptional activity of TCEAL7. (*P < 0.05, **P < 0.01, ***P < 0.001)
Fig 4: TCEAL7 inhibited melanoma cell proliferation, invasion and migration, and induced cell apoptosis. a–f RT-PCR and western blotting assays were used to detect the knockdown and overexpression efficiencies of TCEAL7 in A375 and WM-115 cell lines. g, h CCK-8 assay was used to test cell proliferation following TCEAL7 was overexpressed/silenced in WM-115 and A375 cells. i, j Transwell chamber assay was used to test cell invasion ability following TCEAL7 was overexpressed/silenced in WM-115 and A375 cells. k, l TCEAL7 effect on the apoptosis of WM-115 and A375 cells were detected by flow cytometry. m, n Starch assay was used to test cell migration ability following TCEAL7 was overexpressed/silenced in WM-115 and A375 cells. o, p Western blotting assay was used to measure the expression levels of E-cadherin, N-cadherin and PCNA following TCEAL7 was overexpressed/silenced in WM-115 and A375 cells. (*P < 0.05 and **P < 0.01, compared with Lenti-NC group; #P < 0.05, compared with si-Scramble group)
Fig 5: Analysis of the expression patterns and clinical significance of TCEAL7 in melanoma. a GEPIA database was applied to analyze the expression level of TCEAL7 in melanoma tissues and normal tissues. b, c The expression levels of TCEAL7 in melanoma tissues and nevus tissues at mRNA and protein levels were determined by using RT-PCR and western blotting assays, respectively. d, e. RT-PCR and western blotting were performed to detect the mRNA and protein levels of TCEAL7 in normal melanocyte cell line PIG1 and melanoma cell lines WM-115 and A375. f Kaplan–Meier was used to analyze the prognostic value of TCEAL7 in melanoma patients. (**P < 0.01, ***P < 0.001)
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