Fig 1: Impression cytology and immunofluorescent staining of the samples obtained from the LSCD-affected eye. (a) Central corneal sample. Nuclei are many, well stained with many K7 positive cells (red) with moderate expression (++), and a small number of K-12 positive cells (green) with strong expression (+++). (b) Corneal periphery sample. Nuclei are many, well stained; many K7 positive cells (red) with strong expression (+++), and few K12 positive cells (green) with strong expression (+++). Anti-K7 (OV-TL 12/30) (1:100, ab216016, mouse monoclonal, Abcam, UK); anti-K12 (1:100, ab185627, rabbit monoclonal, Abcam, UK). Confocal laser scanning microscopy, ethanol-fixed samples, immunocytochemistry, and immunofluorescence: green—Alexa Fluor®488 (1:250, ab150077, goat anti-rabbit, Abcam), red—Alexa Fluor®594 (1:250, ab150116, goat anti-mouse, Abcam), blue—Hoechst 33342 (ab228551, Abcam).
Fig 2: Workflow and morphological changes of SH-SY5Y cells following retinoic acid (RA) treatment.(A) Schematic overview of SH-SY5Y cell plating and RA treatment workflow. Undifferentiated SH-SY5Y cells are seeded onto polydopamine (PDA)-coated polydimethylsiloxane (PDMS) wells in complete DMEM, allowed to attach for 24 h, and subsequently cultured in reduced DMEM supplemented with 10 μM RA for at least 3 days. (B) Representative fluorescence micrographs showing SH-SY5Y cells without RA treatment (undifferentiated) and after RA treatment (scale bar: 100 μm). Cells were stained with α-tubulin [primary Ab: Abcam, ab52866; secondary Ab: goat anti-rabbit IgG H&L (Alexa Fluor® 647), Abcam, ab150079, shown in red] to visualize microtubules and Hoechst (Abcam ab228551, shown in blue) to label nuclei. RA-treated cells exhibit elongated, neurite-like extensions consistent with early neuronal differentiation.
Supplier Page from Abcam for Hoechst 33342 Staining Dye Solution