Journal: STAR Protocols

Article Title: Protocol for analysis of miRNAs in human melanoma cells

doi: 10.1016/j.xpro.2025.103861

Figure Lengend Snippet: Spatial localization of hsa-miR-146a-5p, MITF, and BGN in melanocytic and mesenchymal melanoma cells Visualization of hsa-miR-146a-5p (left panel, red), MITF mRNA (middle panel, yellow) and BGN (right panel, green) in melanocytic melanoma cells (M130429, upper row) and in mesenchymal melanoma cells (M130219, lower row). RNAscope stainings were counterstained with DAPI. Images were acquired with the Vectra Polaris. Note that melanocytic cells show higher expression of hsa-miR-146a-5p and MITF , while mesenchymal cells show higher expression of BGN . The scale bar represents 20 μm.

Article Snippet: The subsequent RNAscope Plus smRNA-RNA HD Assay (developed by ACD) is not covered in detail here, as it is fully optimized, and a user manual is available directly from the company.

Techniques: RNAscope, Expressing

Journal: STAR Protocols

Article Title: Protocol for analysis of miRNAs in human melanoma cells

doi: 10.1016/j.xpro.2025.103861

Figure Lengend Snippet:

Article Snippet: The subsequent RNAscope Plus smRNA-RNA HD Assay (developed by ACD) is not covered in detail here, as it is fully optimized, and a user manual is available directly from the company.

Techniques:

Journal: Journal of nanobiotechnology

Article Title: Exosome-derived circUPF2 enhances resistance to targeted therapy by redeploying ferroptosis sensitivity in hepatocellular carcinoma.

doi: 10.1186/s12951-024-02582-6

Figure Lengend Snippet: Fig. 1 Exosomes derived from sorafenib-resistant Huh-7 cells induce an enhanced resistance to sorafenib in HCC cells. (A) Sorafenib-resistant cell line, Huh-7-SR, was established and confirmed by the IC50 value. (B) Co-culture systems were established to investigate the effects of sorafenib-resistant HCC cells on regular HCC cells. The group with additional GW4869 served as a negative control. (C) Cell viabilities of HCC cells co-cultured with Huh-7 or Huh- 7-SR cells under sorafenib stress were evaluated via CCK-8 assays. (D) Assessment of cell proliferation using colony formation assay in Huh-7 and Hep3B cells with the indicated co-culture model. (E) Concentration and size of exosome were measured by NTA. Exosomes were isolated from the supernatant of the culture medium of Huh-7-SR (Exo-SR) and Huh-7 (Exo-Norm) cells by ultracentrifugation. (F) The iconic cup-shaped double-layer closed vesicle structure of isolated exosome was confirmed by TEM. (G) Western blotting analysis of exosomal markers, including CD9, CD63, CD81, LAMP2, TSG101 and Calnexin. An exosome standard from HEK293 cell line was analyzed as a positive control, while Huh-7-SR cell lysate was performed as a negative control. (H) The fluorescent signal of PKH67 labeled Exo-SR was captured in Huh-7 and Hep3B cells. (I) CCK-8 assays were performed to assess the cell viability of Huh-7 and Hep3B cells treated with sorafenib (6 µM) while co-cultured with the indicated exosomes. (J) Colony formation assays were performed to assess the cell proliferation of Huh-7 and Hep3B cells. The data are presented as the mean ± SD of at least three independent experiments. ***P < 0.001

Article Snippet: A positive control was conducted using a standard sample of exosomes derived from HEK293 cells (Novus, USA), while a negative control was performed using Huh-7-SR cell lysate.

Techniques: Derivative Assay, Co-Culture Assay, Negative Control, Cell Culture, CCK-8 Assay, Colony Assay, Concentration Assay, Isolation, Western Blot, Positive Control, Labeling