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. 2014 Apr 21;9(4):e95863. doi: 10.1371/journal.pone.0095863

Figure 2. LAMTOR2 knockdown increases neurite formation of NGF- but not EGF-treated PC12 cells.

Figure 2

(A, B) PC12 cells, transfected either with control or specific LAMTOR2 siRNA, were left untreated or stimulated with NGF (25 ng/ml) or EGF (100 ng/ml). Neurite outgrowth was examined in 3–5 independent microscopic fields (72 h after stimulation shown in (A)). Percentages of neurite-bearing cells after various time points (6 h –72 h) were calculated and are depicted in (B). Microscopy images are representative of 3 independent experiments. Values represent the mean ± SEM, n = 3. Scale bar = 20 µm. Differences were analyzed using unpaired two-tailed t-test: *p<0.05. (C) Analyses of neurite length in µm. Values represent the mean ± SEM, n = 9. Differences were analyzed using unpaired two-tailed t-test: *p<0.05. (R1-1.2a). (D) The expression of the plasticity protein GAP-43 was analyzed in control- (i, iii and v) and LAMTOR2 knockdown (ii, iv and vi) PC12 cells that were stimulated with (iii–vi) and without NGF (unstimulated control i-ii) in the presence (v-vi) and absence (i-iv) of the TrkA inhibitor K-252a. Icons in iv (higher magnification of GAP-43 stained neurites). Microscopy images are representative of 4 independent experiments. Scale bar = 10 µm.