Journal: Frontiers in Chemistry
Article Title: An imidazole functionalized pentameric thiophene displays different staining patterns in normal and malignant cells
doi: 10.3389/fchem.2015.00058
Figure Lengend Snippet: Intracellular targets of p-HTIm . Merged fluorescence images of (A,C) human fibroblasts (AG01518) and (B,D) melanoma cells (SK-MEL-28) stained with p-HTIm (5 μM, 30 min; green) and incubated in fresh medium for 24 h. The cells were stained with fluorescent markers against α-tubulin, autophagosomes (LC3), ER (calnexin), fibronectin, p62, peroxisomes (PMP70), proteasome 20S, PTEN, recycling endosomes (rab11a), Golgi (Golga2), mitochondria (mitotracker), and lysosomes (LAMP-2) (red). Cell nuclei were labeled with DAPI (blue). The color channels are split in ( C ) and ( D ) to improve the visualization. Scale bars 20 μm.
Article Snippet: For immunostaining, PTAA-stained cells were after fixation permeabilized with 0.1% saponin (Sigma-Aldrich) in PBS containing 5% fetal bovine serum (20 min, room temperature) and incubated for 2 h at room temperature with one of the following monoclonal mouse primary antibodies: Golga2/GM130 (1:250, Novus Biologicals, Littleton, CO, USA), lysosome-associated membrane protein 2 (LAMP-2, 1:100; Southern Biotech, Birmingham, AL, USA), p62 (1:100, BD Biosciences, Franklin Lakes, NJ, USA), or polyclonal anti-rabbit primary antibodies; α-tubulin (1:1000, Abcam, Cambridge, UK), calnexin (1:700, Novus Biologicals), early endosomal antigen-1 (EEA-1, 1:400; Sigma-Aldrich), fibronectin (1:400, Sigma-Aldrich), LC3B (1:100, Novus Biologicals), Niemann-Pick type C1 (NPC1, 1:250; Abcam), peroxisomal membrane protein 70 (PMP70, 1:1000; Molecular Probes), PTEN (1:20, Novus Biologicals), anti-Rab11a (1:200, Abcam), and proteasome 20S (1:100, Abcam).
Techniques: Fluorescence, Staining, Incubation, Labeling
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