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

Figure 2.

Figure 2.

3-MA, and Con A treatments inhibited NF protein degradation. (A) Treatment of NF-gene-transfected N2a cells with 3-MA (5 mM, 42 h) and measurement of levels of NF proteins (NEFL, 3-fold, p = 0.0003; NEFM, 2.3-fold, p < 0.0015 and NEFH-MYC, 1.4-fold, p < 0.0042; n = 3 for all subunits). NF-gene-transfected N2a cells (NFs) were treated with 3-MA (NF + 3-MA), stained with NEFL and NEFM antibodies (B, lower and higher mag. Scale bars: 10-µm) and quantification of NEFL (C, n = 115 cells for NF and n = 122 cells for NF + 3-MA, p < 0.0001) and NEFM (D, n = 119 cells for NF and n = 110 for NF + 3-MA, p < 0.0001, unpaired t-test) staining in cells. Con A treatment of NF-gene-transfected cells and measurement of levels of NF proteins (E, NEFL, 2-fold, n = 4, p < 0.0051; NEFM, 2-fold, n = 4, p < 0.0188; NEFH-MYC, 3-fold, n = 3, p < 0.0116), MAP1LC3-I (E, n = 4, p < 0.0054), MAP1LC3-II (E, n = 4, p < 0.0003), the ratio of MAP1LC3-II:MAP1LC3-I (E, n = 4, p < 0.0001), and the formation of mature-CTSD (E, n = 4, p < 0.0091). Panels A-E, unpaired t-test; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.