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. 2021 Feb 5;11(2):222. doi: 10.3390/biom11020222

Figure 1.

Figure 1

Cardiolipin translocates to MAM subdomains during autophagy. (A) Isolated MAMs and pure mitochondria fractions obtained from human 2FTGH (2F) fibroblasts cells, untreated or treated with HBSS for 1 h at 37 °C were subjected to phospholipid extraction. All fractions were run on HPLC plates and analyzed by HPTLC immunostaining for the presence of CL using anti-CL as described in Materials and Methods. A positive control was obtained using pure standard CL (STD, Sigma-Aldrich). (B) The purity of MAMs and pure mitochondria fractions was also tested by Western blotting for the presence of specific markers: CANX for MAMs or VDAC1 for mitochondria and MAMs, Cytc for mitochondria. (C) Autophagy was checked by Western blot analysis, using rabbit anti-LC3 PAb or rabbit anti-p62/SQSTM1 pAb. Loading control was evaluated using anti-ACTB Mab. A representative experiment among 3 is shown. Bar graph on the right shows densitometric analysis. Results represent the mean ± SD from 3 independent experiments. *** p ≤ 0.001, HBSS vs. control cells.