Journal: Frontiers in Immunology

Article Title: Establishment and validation of evaluation models for post-inflammatory pigmentation abnormalities

doi: 10.3389/fimmu.2022.991594

Figure Lengend Snippet: The role of IL-37 in melanogenesis. MNT1 cells were treated with different concentrations of IL-37. (A) qRT-PCR was used to examine the mRNA levels of melanogenesis-related genes. (B) The protein levels of MITF, TYR, TYRP1, DCT, PMEL17 and MLANA were analyzed by Western blotting. (C) Tyrosinase activity was measured by L-DOPA assay. (D) representative immunofluorescence images showing the distribution of melanosomes. (N = 3, ANOVA, error bar represents mean ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: Primary antibodies specific for GAPDH (#AP0066, Bioworld), TYR (BS1484, Bioworld), MITF (STJ94134, St. John’s Laboratory), TYRP1 (ab235447, Abcam), DCT (NBP1-56058, Novusbio), PMEL17(#H1219, Santa Cruz and bs-17478R, Bioss) and MLANA (bs-0051R, Bioss and ET1610-47, HUABIO) were purchased as indicated.

Techniques: Quantitative RT-PCR, Western Blot, Activity Assay, Immunofluorescence

Journal: bioRxiv

Article Title: Mitochondrial Signatures Shape Phenotype Switching and Apoptosis in Response to PLK1 and RSK Inhibitors in Melanoma

doi: 10.1101/2024.06.14.599035

Figure Lengend Snippet: (A) CXCL8 transcript levels in MeWo and A-375 cells treated for 72 h with DMSO 0.1 % (v/v) or a panel of inhibitors targeting putative BI-D1870 targets. NA: non-available due to cell death. (B) Viable cell counts of melanoma cells with Presto Blue HS. (C) Representative crystal violet staining of cells treated as in A. (D) Quantification of crystal violet staining, as in C. (E,F) Immunoblotting of cell extracts from cells treated as indicated for 72 h. (G) Densitometric analysis of immunoblotting from 3 independent experiments related to E and F. (H,I) PLK1 and CXCL8 transcript levels in cells 72 h after PLK1 knockdown. (J) Immunoblotting of cell extracts from cells treated as in H-I. Numbers indicate independent experiments. (K) Densitometric analysis of immunoblotting from 3 independent experiments related to J. All data are shown as means ± SEM from 3 independent experiments. A: * p < 0.05, unpaired Student’s t -test (cell lines tested separately). D,K: * p < 0.05, paired Student’s t -test. G-I: * p < 0.05, Two-Way ANOVA with Tukey multiple comparison test.

Article Snippet: Primary antibodies against PARP (#9542), β -actin (#3700), PLK1 (#4535), and Melan-A/MART-1/MLANA (#64718) were purchased from Cell Signaling Technology Inc. (Whitby, ON, CA).

Techniques: Staining, Western Blot, Knockdown, Comparison

Journal: bioRxiv

Article Title: Mitochondrial Signatures Shape Phenotype Switching and Apoptosis in Response to PLK1 and RSK Inhibitors in Melanoma

doi: 10.1101/2024.06.14.599035

Figure Lengend Snippet: (A, B, C) Gene expression in MeWo and A-375 cells treated for 72 h as indicated. NA: Not available due to cell death. (D) Immunoblotting of protein extracts from MeWo cells treated as in A-C. Numbers indicate independent experiments. (E) Gene expression of differentiation markers in cells treated with siRNA against PLK1 versus non-targeting siRNA for 72 h. (F) Heat-map of Log2 fold change of specific transcripts linked to phenotype switching in MeWo cells treated with BI-D1870 (3 µM) for 72 h. Derived from RNA-Seq analyses in . (G,H) Gene expression in MeWo cells treated with BI 6727 (1 µM) or BRD7389 (10 µM) for 72 h. All data are shown as mean ± SEM from 3 independent experiments. A-C, E: * p < 0.05, Two-Way ANOVA with Tukey multiple comparison test. F: * p-adj < 0.05. G-H: * < 0.05, paired Student’s t -test.

Article Snippet: Primary antibodies against PARP (#9542), β -actin (#3700), PLK1 (#4535), and Melan-A/MART-1/MLANA (#64718) were purchased from Cell Signaling Technology Inc. (Whitby, ON, CA).

Techniques: Gene Expression, Western Blot, Derivative Assay, RNA Sequencing, Comparison

Journal: bioRxiv

Article Title: Mitochondrial Signatures Shape Phenotype Switching and Apoptosis in Response to PLK1 and RSK Inhibitors in Melanoma

doi: 10.1101/2024.06.14.599035

Figure Lengend Snippet: (A) Schematic depicting the strategy to identify transcripts associated with resistance to PLK1 targeting. The six PLK inhibitors, in addition to BI-D1870, are indicated in panel E. (B) Transcripts that significantly correlate (p < 0.05) with 2 or more PLK1-directed treatments were selected for further analyses. (C) Functional gene analysis (g:Profiler, GO:BP) performed on correlating transcripts from B. Bars indicate p-adj values and circles indicate number of transcripts associated with a gene set. (D) Essentiality of transcripts encoding for mitochondrial proteins that significantly correlate with resistance to PLK targeting in melanoma cells (DepMap). (E) Heat-map representation and hierarchical clustering of Spearman correlations between the 124 mitochondrial signature transcripts and respective impact on proliferation dynamics of BI-D1870 and PLK1 targeting approaches (DepMap). (F) Functional classification of 124 mitochondrial signature transcripts associated with resistance to PLK1 targeting, according to MitoPathways3.0. (G) STRING interaction network of the 124 mitochondrial signature transcripts associated with resistance to PLK1 targeting. Nodes indicate proteins, and edges indicate interactions. Spearman’s correlation with proliferation dynamics (DepMap) is indicated by nodes’ colored borders. (H) Gene expression in MeWo cells transfected with the indicated siRNA for 48 h and treated with DMSO 0.1 % (v/v) or BI 6726 (100 nM) for 72 h. (I) Relative viable cell counts of MeWo cells treated as in H but with 10-fold dilutions of BI 6727, using Presto Blue HS. (J) Model depicting the impact of mitochondrial signatures on the response to RSK and PLK inhibitors. All data are shown as means ± SEM from 3 or more independent experiments. * p < 0.05, One-Way ANOVA with Dunnett multiple comparison test.

Article Snippet: Primary antibodies against PARP (#9542), β -actin (#3700), PLK1 (#4535), and Melan-A/MART-1/MLANA (#64718) were purchased from Cell Signaling Technology Inc. (Whitby, ON, CA).

Techniques: Functional Assay, Gene Expression, Transfection, Comparison

Journal:

Article Title: ALTERED EXPRESSION OF THE IRON TRANSPORTER Nramp1 (Slc11a1) DURING FETAL DEVELOPMENT OF THE RETINAL PIGMENT EPITHELIUM IN MICROPHTHALMIA-ASSOCIATED TRANSCRIPTION FACTOR Mitf mi and Mitf vitiligo MOUSE MUTANTS

doi: 10.1016/j.exer.2007.11.015

Figure Lengend Snippet: Comparison of Affymetrix Mouse 430_2 Microarray Results

Article Snippet: Mlana , Melan-A (MART1) , Mm01195780_m1 , GOI , Forms a complex with Silver (Pmol17); critical role in expression, stability, trafficking, and processing of Silver.

Techniques: Comparison, Microarray, Binding Assay

Journal:

Article Title: ALTERED EXPRESSION OF THE IRON TRANSPORTER Nramp1 (Slc11a1) DURING FETAL DEVELOPMENT OF THE RETINAL PIGMENT EPITHELIUM IN MICROPHTHALMIA-ASSOCIATED TRANSCRIPTION FACTOR Mitf mi and Mitf vitiligo MOUSE MUTANTS

doi: 10.1016/j.exer.2007.11.015

Figure Lengend Snippet: IRON HQMEOSTASIS, PJGMENT FORMATION AND REDOX QRT-PCR PRIMER/PROBE LIST

Article Snippet: Mlana , Melan-A (MART1) , Mm01195780_m1 , GOI , Forms a complex with Silver (Pmol17); critical role in expression, stability, trafficking, and processing of Silver.

Techniques: Expressing, Activity Assay, Disruption, Sequencing, Membrane, Dispersion, Ubiquitin Proteomics

Journal:

Article Title: ALTERED EXPRESSION OF THE IRON TRANSPORTER Nramp1 (Slc11a1) DURING FETAL DEVELOPMENT OF THE RETINAL PIGMENT EPITHELIUM IN MICROPHTHALMIA-ASSOCIATED TRANSCRIPTION FACTOR Mitf mi and Mitf vitiligo MOUSE MUTANTS

doi: 10.1016/j.exer.2007.11.015

Figure Lengend Snippet: Quantitative RT-PCR expression analysis of additional genes involved in iron homeostasis demonstrates significant downregulation of the Transferrin Receptor (Tfrc), which is involved in iron uptake into the cell, and conversely, significant upregulation of Ceruloplasmin (Cp), and Slc40a1 (Ferroportin), which are involved in iron export from the cell. Epoxide hydrolase (Ephx1) was significantly upregulated in Mitfvit/vit mutants relatived to C57BL/6 controls, while the pigment-related genes Gpnmb and Mlana were significantly downregulated (*p<.05, **p<.001). Numbers above 1.00 on the y-axis represent the relative fold-change increase or upregulation in gene expression, while numbers below 1.00 on the y-axis represent the relative fold-change decrease or downregulation in gene expression in Mitfvit/vit mutants relative to C57BL/6 controls. Ephx1 (epoxide hydrolase); Slc11a1 (Nramp1); Slc40a1 (Ferroportin); Cp (Ceruloplasmin); Fxn (Frataxin); Heph (Hephaestin); Si (Silver; Pmel17); Trf (Transferrin); Fth1 (Ferritin, heavy chain); Slc11a2 (Nramp2); Trfc (Transferrin Receptor); Gpnmb (Glycoprotein, membrane bound); Mlana (MART-1; Melanoma antigen).

Article Snippet: Mlana , Melan-A (MART1) , Mm01195780_m1 , GOI , Forms a complex with Silver (Pmol17); critical role in expression, stability, trafficking, and processing of Silver.

Techniques: Quantitative RT-PCR, Expressing, Gene Expression, Membrane