Immunofluorescence Staining of Fibroblasts

Fibroblasts were seeded in 48-well plates at 6060 cells/cm² and, after overnight attachment, cells were treated with drugs as described above. After 1 and 4 d, cells were fixed with paraformaldehyde and stained for immunofluorescence using primary mouse anti-α-SMA (Santa Cruz Biotechnology, Dallas, TX, USA, sc-32251) and secondary goat anti-mouse IgG AlexaFluor488-conjugated antibodies (Thermo Fisher Scientific A-11001), and DNA was counterstained with Hoechst 33342 as previously described [50 (link)].

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Weathers P., Towler M., Kiani B.H., Dolivo D, & Dominko T. (2024). Differential Anti-Fibrotic and Remodeling Responses of Human Dermal Fibroblasts to Artemisia sp., Artemisinin, and Its Derivatives. Molecules, 29(9), 2107.

Publication 2024

mouse anti-α-SMA

Corresponding Organization : Worcester Polytechnic Institute

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Variable analysis

independent variables

Drugs (treatments)

dependent variables

α-SMA expression (as measured by immunofluorescence)

control variables

Fibroblast seeding density (6060 cells/cm^2)
Attachment time (overnight)
Staining procedure (paraformaldehyde fixation, primary and secondary antibodies, Hoechst 33342 DNA counterstain)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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