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Figure 4 | BMC Neuroscience

Figure 4

From: Upregulation of myeloid cell leukemia-1 potentially modulates beclin-1-dependent autophagy in ischemic stroke in rats

Figure 4

Mcl1 potentially modulated beclin-1-induced autophagy. (A) Double immunostaining showed the co-localization of beclin-1 and Mcl1 in brains from control (a) and MCAO group (4 h, 24 h, 72 h, b-d). (B) Quantitative assessment the numbers of beclin-1+, Mcl1+ and beclin-1+/caspase-3+ cells in control and MCAO group. Results were expressed as mean ± SD from four independent experiments. *P < 0.001 vs. control group, #P < 0.001 vs. the previous time point. (C) Some beclin-1expressing cells displayed the characteristic features of apoptosis including cell shrinkage and pyknotic nuclei (small arrow), whereas those cells coexpressing Mcl1 and beclin-1 displayed little chromatin clumping, and their nuclei were not pyknotic, as demonstrated by co-labeling with DAPI (big arrows). (D) Pretreatment with 3-MA before MCAO significantly blocked autophagy but did not affect the protein levels of Mcl1. Western blot results showed the levels of LC3-II significantly decreased in 3-MA group compared with that of NS group (a). However, Mcl1 protein levels remained unchanged in 3-MA group compared with that of NS group after autophagy inhibition (b). Optical densities of respective protein bands were analyzed with Image J 1.42q and normalized to the loading control (β-actin). Results were expressed as mean ± SD from four independent experiments. *P < 0.001 vs. control group; #P < 0.001, &P > 0.10 vs. NS group. Cont: control group. Scale bars = 50 μm (in A), 20 μm (in C).

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