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. 2002 Jan;13(1):12–24. doi: 10.1091/mbc.01-04-0210

Figure 4.

Figure 4

Isolated actin-binding site from PKCε inhibits neurite outgrowth. (A) Expression vectors encoding the actin-binding site from PKCε, or a scrambled version of this site, fused to either EGFP or a myc-tag via a linker sequence was constructed. (B) SK-N-BE(2) cells were cotransfected in a 1:7 ratio with vectors encoding EGFP, εFL-EGFP, or εRD-EGFP and myc-tagged ABS (ABS), scrambled ABS (Scram), or empty myc-vector (−). Cells were fixed 16 h after transfection and transfected cells, identified with EGFP fluorescence, were counted and the number of cells with long neurites was determined. Data (mean ± SEM, n = 3) are presented as percentage of transfected cells with long neurites. EGFP-tagged ABS, scrambled actin-binding site (Scram), and EGFP alone (EGFP) were expressed in SK-N-BE(2) cells (C) and SH-SY5Y/TrkA cells (D). The cells were incubated in regular medium (−) or treated with 10 μM RA for 2 d or with 100 ng/ml NGF for 4 d (RA, NGF). The cells were fixed and the number of transfected cells bearing long neurites was assessed. Data (mean ± SEM, n = 3) are presented as percentage of transfected cells with long neurites. ∗, statistically significant differences with analysis of variance followed by Duncan's multiple range test compared with similar conditions by using control vector instead of ABS vector.