Mitofusin 2 Promotes Apoptosis of CD4⁺ T Cells by Inhibiting Autophagy in Sepsis

Figure 2

LPS stimulation increased Mfn2 expression, downregulated autophagy, and elevated apoptosis in murine splenic CD4⁺ T cells. Murine splenic CD4⁺T cells were isolated and subjected to in vitro culture with LPS (10, 100, and 1000 ng/ml) or PBS for 24 hours. (a) Representative immunoblots and densitometric values of Mfn2. β-Actin served as an internal control. (b) Representative dot plots and the percentage of Annexin-V-positive cells. (c) Representative histograms and the percentage of LC3-II-positive cells. (d) Whole cell lysate was used for immunoblot analysis of Beclin1, p62, and LC3-II expression. β-Actin served as an internal control for Beclin1 and p62 protein expression levels. (e) Representative images of immunofluorescence staining. DAPI and Cy3 were used to label nuclei and endogenous LC3, respectively. The arrows indicate LC3 puncta. Quantification of the average number of LC3 puncta per cell ( well, 3 independent experiments, and >50 cells examined per experiment). Data from three independent experiments are shown as the mean ± SD. Bar 1, the LPS 0 group; bar 2, the LPS 10 group; bar 3, the LPS 100 group; bar 4, the LPS 1000 group. , significant difference versus the LPS 0 group. , significant difference versus the LPS 0 group. ^#, significant difference versus the LPS 10 group. ^##, significant difference versus the LPS 10 group. ^$, significant difference versus the LPS 100 group. ^$$, significant difference versus the LPS 100 group.

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