<p>(<b>A</b>) Undifferentiated and retinoic-acid (RA) differentiated SHSY-5Y cells were lysed and levels of total and phosphorylated-cofilin were assayed using Western blot analysis with specific antibodies. Representative blots are shown. GAPDH was used as an additional loading control. Levels of p-cofilin were quantified from 2 independent experiments. The p-cofilin/cofilin ratio was set as 1 in mock treated cells. (<b>B</b>) RA-differentiated SHSY-5Y cells were additionally treated with HoPan or HoPan and pantethine. Levels of p-cofilin were assayed and quantified as in A. (<b>C</b>) Control SHSY-5Y cells were differentiated with RA, plated on poly-lysine and analyzed after 48 hours for their morphology (using rhodamin-phalloidin marking F-actin, red) and differentiation status (using β-III-Tubulin staining, green). In RA-treated cultures undifferentiated and differentiated cells are visible. Undifferentiated cells showed very short filopodia and did not express β-III-Tubulin (indicated as arrowheads). 48 hours after seeding many differentiated cells showed an elongated shape, neurite growth and high expression of β-III-Tubulin (indicated as arrows). (<b>D–F</b>) Control (D), HoPan (E) or HoPan plus pantethine (F) treated cells were differentiated with RA and plated on poly-lysine. 48 hours after seeding cells were fixed and stained for β-III-tubulin, representative parts of cell morphology and β-III-Tubulin staining of the different conditions are shown. The percentage of neurite-bearing cells was calculated for each condition. Scale bars represent 25 µm (<b>C–F</b>). (<b>G</b>) Quantification of cells showing β-III-Tubulin positive neurites is presented.</p
