Cells seeded on Lab-Tek®II Chamber SlideTM system (Thermo Fisher Scientific Inc., Rochester, NY, USA) were fixed with PLP solution for 45 minutes61 (link).
For immunocytochemical staining, endogenous peroxidase was blocked with 3% H2O2 and cells were pre-incubated with serum14 (link), 20 (link). Primary antibody against α-SMA (Abcam) was used. After washing, cells were incubated with secondary antibody (biotinylated IgG; Sigma), washed, and processed using Vectastain ABC-HRP kits (Vector Laboratories Inc.). Cells were counterstained with hematoxylin and mounted with Biomount medium on slides as previously described14 (link), 20 (link).
For immunofluorescence, cells were pre-incubated with serum. Primary antibodies against α-SMA (Abcam) and affinity-purified Alexa Fluor 488-conjugated secondary antibody (Life Technologies Corporation, USA) were used. Nuclei were labeled with DAPI (Sigma-Aldrich, USA) and mounted with medium to prevent quenching (Vector Laboratories Inc., USA)61 (link).
For immunocytochemical staining, endogenous peroxidase was blocked with 3% H2O2 and cells were pre-incubated with serum14 (link), 20 (link). Primary antibody against α-SMA (Abcam) was used. After washing, cells were incubated with secondary antibody (biotinylated IgG; Sigma), washed, and processed using Vectastain ABC-HRP kits (Vector Laboratories Inc.). Cells were counterstained with hematoxylin and mounted with Biomount medium on slides as previously described14 (link), 20 (link).
For immunofluorescence, cells were pre-incubated with serum. Primary antibodies against α-SMA (Abcam) and affinity-purified Alexa Fluor 488-conjugated secondary antibody (Life Technologies Corporation, USA) were used. Nuclei were labeled with DAPI (Sigma-Aldrich, USA) and mounted with medium to prevent quenching (Vector Laboratories Inc., USA)61 (link).
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