Western blot analysis
Proteins from spleen tissues were separated by SDS-PAGE and transferred
to PVDF membranes. The membranes were then incubated overnight at 4 °C
with antibodies against the protein of interest, including BCL-2 (D17C4;
3498; Cell signaling), Bax (D2E11; 5023; Cell signaling), GAPDH (D16H11;
5174T; Cell signaling) over-night at 4 °C. Protein quantification was
measured in optical density units using Image Lab software (Bio-Rad, CA,
USA) and was normalized to the corresponding sample expression of GAPDH.