Figure 3.

miR-296-5p overexpression enhances lymphocyte apoptosis by targeting Bcl-xl. (A) The expression of pro-apoptotic and anti-apoptotic proteins normalized to GAPDH in PBMCs of patients with T1DM determined by Western blot assay (n = 30). (B) Quantitative analysis of apoptosis of PBMCs in patients with T1DM compared with healthy individuals analyzed using flow cytometry. (C) The protein expression of BCL-XL and C-caspase3 normalized to GAPDH determined by Western blot assay after lymphocytes transfected with vector and pCMV-Bcl-xl. (D) Quantitative analysis of apoptosis of lymphocytes transfected with vector and pCMV-Bcl-xl analyzed using flow cytometry. (E) The protein expression of BCL-XL and C-caspase3 normalized to GAPDH determined by Western blot assay after lymphocytes transfected with miR-296-5p inhibitor or inhibitor NC. (F) Quantitative analysis of apoptosis of lymphocytes after transfected with miR-296-5p inhibitor or inhibitor NC evaluated by flow cytometry. (G) Rescue experiments on the protein expression of BCL-XL and C-caspase3 normalized to GAPDH determined by Western blot assay. (H) Rescue experiments on the apoptosis of lymphocytes evaluated by flow cytometry. ^*p < 0.05 compared with cells transfected with control, vector, or inhibitor NC, ^#p < 0.05 compared with cells transfected with miR-296 inhibitor + si-NC. The results were measurement data and expressed as mean ± standard deviation. Data comparison between two groups was performed using unpaired sample t-test, and data comparisons among multiple groups were performed using one-way analysis of variance. The cell experiment was repeated three times independently.