Journal: Scientific Reports

Article Title: Nuclear Transport Factor 2 (NTF2) suppresses WM983B metastatic melanoma by modifying cell migration, metastasis, and gene expression

doi: 10.1038/s41598-021-02803-0

Figure Lengend Snippet: NTF2 affects lamin A nuclear localization, nuclear size, and heterochromatin distribution. VGP primary melanoma refers to a primary vertical growth phase melanoma cell line (WM983A) that was derived from the same patient as NTF2 low (WM983B) and represents an earlier stage of disease. (A) The indicated cells were stained for lamin A and representative images are shown. Images were acquired with the same exposure time and brightness/contrast were identically adjusted. Line scans were acquired across the nuclear rim as indicated by the yellow line. In the top graph, lamin A intensity was quantified across at least 50 line scans per cell line for two independent experiments (see “ ”). To measure the lamin A intensity at the nuclear envelope (NE), the highest lamin A intensities from each line scan were averaged. To measure the total nuclear lamin A intensity, the total fluorescence intensity of thresholded nuclei was quantified for at least 102 nuclei per condition. To measure the NE/nucleoplasm lamin A intensity, the maximum value along the line scan was divided by the minimum value within the nucleus. Error bars are SD. One-way ANOVA followed by Dunnett’s multiple comparisons test with ***p < 0.0001 and ns not significant. We also measured a 44% reduction in lamin B1 staining intensity at the NE in NTF2 high dox + cells compared to NTF2 low cells (data not shown). (B) Nuclear cross-sectional area, which serves as a good proxy for nuclear surface area and volume , , , , was measured for at least 274 Hoechst-stained nuclei per cell line (918 nuclei on average). The normal melanocyte cell line is AG21857. Error bars are SD. One-way ANOVA followed by Dunnett’s multiple comparisons test with ***p < 0.0001. (C) The indicated cells were stained for the heterochromatin marker H3K9me3 and representative images are shown. Images were acquired with the same exposure time and brightness/contrast were identically adjusted. To quantify heterochromatin distribution, H3K9me3 intensity was quantified along line scans drawn from the nuclear rim toward the center (see “ ”). Averages from at least 150 line scans are shown for each cell line for two independent experiments. Error bars are SEM. One-way ANOVA followed by Dunnett’s multiple comparisons test with **p < 0.01, ***p < 0.0001. The statistical comparisons are for NTF2 low and VGP primary melanoma compared to NTF2 high dox + at line scan position 0.2 µm.

Article Snippet: The human melanocyte cell line (AG21857) was obtained from the NIA Cell Culture Repository through the Coriell Institute (only used in Fig. B).

Techniques: Derivative Assay, Staining, Fluorescence, Marker

Journal: Heliyon

Article Title: Identification of the role of autophagy-related TNFSF10/ hsa-let-7a-5p axis in vitiligo development and potential herbs exploring based on a bioinformatics analysis

doi: 10.1016/j.heliyon.2023.e23220

Figure Lengend Snippet: Effects of EGCG on the 3-MA-treated PIG1 melanocytes. (A) The percentages of viable cells change in PIG1 after treatment with varying concentrations of 3-MA (0–10 mM). (B) The cell viability of PIG1 treated with 3-MA and 3-MA + EGCG by the CCK-8 assay. (C) Real-time PCR of changes in mRNA levels of TNFSF10，MAPK1 and BID in PIG1. Data were presented as mean ± SEM of three independent experiments; **P < 0.01 compared with the control group; ##P < 0.01 compared with the 3-MA group.

Article Snippet: The human melanocyte line PIG1 was purchased from ScienCell Research Laboratories, Inc (California, USA).

Techniques: CCK-8 Assay, Real-time Polymerase Chain Reaction, Control

Journal: Experimental and Therapeutic Medicine

Article Title: ZMIZ1 promotes the proliferation and migration of melanocytes in vitiligo

doi: 10.3892/etm.2020.8849

Figure Lengend Snippet: Effects of ZMIZ1 on the proliferation and apoptosis of PIG1 and PIG3V cells. (A) Effect of ZMIZ1 on the proliferation of PIG1 and PIG3V cells was analyzed using an MTT assay. (B) Effect of ZMIZ1 on the apoptosis of PIG1 and PIG3V cells was analyzed using flow cytometry. * P<0.05, ** P<0.01 and *** P<0.001 vs. Con. ## P<0.01 and ### P<0.001 vs. OE NC. Con, control; sh, short hairpin RNA; OE, overexpression; NC, negative control; ZMIZ1, zinc finger MIZ-type containing 1; OD, optical density.

Article Snippet: The human normal melanocyte cell line PIG1 and human vitiligo melanocyte cell line PIG3V were purchased from ScienCell Research Laboratories, Inc.

Techniques: MTT Assay, Flow Cytometry, Control, shRNA, Over Expression, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: ZMIZ1 promotes the proliferation and migration of melanocytes in vitiligo

doi: 10.3892/etm.2020.8849

Figure Lengend Snippet: ZMIZ1 regulates the expression levels of Bcl-2 and caspase-3 in PIG1 and PIG3V cells. Effect of ZMIZ1 on the expression levels of proliferation- and apoptosis-related proteins in PIG1 and PIG3V cells was analyzed using western blotting. * P<0.05, ** P<0.01 and *** P<0.001 vs. Con. ## P<0.01 vs. OE NC. Con, control; sh, short hairpin RNA; OE, overexpression; NC, negative control; ZMIZ1, zinc finger MIZ-type containing 1; OD, optical density.

Article Snippet: The human normal melanocyte cell line PIG1 and human vitiligo melanocyte cell line PIG3V were purchased from ScienCell Research Laboratories, Inc.

Techniques: Expressing, Western Blot, Control, shRNA, Over Expression, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: ZMIZ1 promotes the proliferation and migration of melanocytes in vitiligo

doi: 10.3892/etm.2020.8849

Figure Lengend Snippet: ZMIZ1 regulates the migration of PIG1 and PIG3V melanocytes. Effect of ZMIZ1 on the migration of PIG1 and PIG3V cells was analyzed using a Transwell assay. Magnification, x200. * P<0.05 and ** P<0.01 vs. Con. # P<0.05 vs. OE NC. Con, control; sh, short hairpin RNA; OE, overexpression; NC, negative control; ZMIZ1, zinc finger MIZ-type containing 1; OD, optical density.

Article Snippet: The human normal melanocyte cell line PIG1 and human vitiligo melanocyte cell line PIG3V were purchased from ScienCell Research Laboratories, Inc.

Techniques: Migration, Transwell Assay, Control, shRNA, Over Expression, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: ZMIZ1 promotes the proliferation and migration of melanocytes in vitiligo

doi: 10.3892/etm.2020.8849

Figure Lengend Snippet: Effect of ZMIZ1 on the expression levels of F-actin cytoskeleton protein in PIG1 and PIG3V cells in vitro was analyzed using immunocytochemistry staining. Magnification, x63. Con, control; sh, short hairpin RNA; OE, overexpression; NC, negative control; ZMIZ1, zinc finger MIZ-type containing 1; OD, optical density.

Article Snippet: The human normal melanocyte cell line PIG1 and human vitiligo melanocyte cell line PIG3V were purchased from ScienCell Research Laboratories, Inc.

Techniques: Expressing, In Vitro, Immunocytochemistry, Staining, Control, shRNA, Over Expression, Negative Control