In Vivo Suppression of Autophagy via Lentiviral shRNA Targeting Atg5 Improves Lupus-Like Syndrome

Figure 5

Silencing Atg5 reduced the number of several immune cells in the lymph nodes of Trem-1^-/-.lpr mice. shLuc- or shAtg5-lentivirus-infected mice were euthanized at 38-40 weeks old. Single-cell suspension from the lymph nodes was obtained by passing them through a nylon filter. (a) Single-cell suspension was stained with AO and analyzed using flow cytometry to determine the autophagy level (AVOs (%)). (b) Total lymph node cell numbers were counted and compared. (c, d, e) The percentages (left) and cell numbers (right) of different immune cell subsets were evaluated by flow cytometry using various cell-specific markers, including total B cell (CD19⁺), total T cell (CD3⁺), mDC (CD11c⁺B220^-), pDC (CD11c⁺B220⁺), T1+T2 B (transitional B cell, CD19⁺CD21^-/lowCD23^-/low), FoB (follicular B cell, CD19⁺CD21⁺CD23^hi), MZB (marginal zone B cell, CD19⁺CD21^hiCD23⁺), PC (plasma cell, CD138⁺), DN T (double-negative T cell, CD3⁺CD4^-CD8^-, DP T (double-positive T cell, CD3⁺CD4⁺CD8⁺), T helper cell (CD3⁺CD4⁺), cytotoxic T cell (CD3⁺CD8⁺), and regulatory T cell (CD3⁺CD4⁺CD25⁺FoxP3⁺). Mean values were shown by a bar.

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