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. 2022 Sep 21;19(4):1277–1292. doi: 10.1080/15548627.2022.2124500

Figure 1.

Figure 1.

NF formation and degradation in N2a cells. Untransfected (Mock, A) and NF-gene-transfected (NFs, B) N2a cells were immunolabeled with NEFL and NEFM antibodies (A and B). Scale bar: 10 µm. Electron microscopy of NF containing cells show formation of parallel NF filaments (purple arrows) that are interconnected with cross-bridges (C, see red arrows). Immunolabeling of these filamentous cells with NF antibodies (D, NEFL, 10 nm and NEFHCOOH, 6 nm) (E, NEFM, 10 nm and NEFL, 6 nm gold particles) show labeling of filaments with all 3 NF antibodies (NEFL, NEFM and NEFH). Scale bar: 50 nm. NF-transfected cells treated with cycloheximide (CHX) show reduction in NF proteins at 1.5 h (F and G, compared to 100% at 0 h time point, NEFL 70%, n = 4, p = 0.0115; NEFM 79%, n = 4, p = 0.0439; NEFH-MYC-79%, n = 4, p = 0.0240), and at 3 h time point (F and G, compared to 100% at 0 h time point, NEFL 53%, n = 5, p = 0.0001; NEFM 69%, n = 5, p = 0.0113; NEFH-MYC 77%, n = 8, p = 0.0007; n = 8). *p < 0.05; ***p < 0.001, one-way ANOVA.