File S1 - Necroptosis Takes Place in Human Immunodeficiency Virus Type-1 (HIV-1)-Infected CD4+ T Lymphocytes

Supporting Figures. Figure S1 - Uninfected cells grow with a low percentage of apoptosis and necrosis. Various cells were cultured in conditioned RPMI 1640 medium. After 4 days, cells were harvested and analyzed by flow cytometry together with HIV-1-infected cells. Data in this figure is representative of at least 3 independent experiments. Figure S2 - Nec-1 has no effect on uninfected cells. The primary CD4+T cells were cultured in conditioned RPMI 1640 medium and cells were treated with DMSO or Nec-1. After 4 days, cells were harvested and analyzed by flow cytometry. Data in this figure is representative of at least 3 independent experiments. Figure S3 - Nec-1 or NSA inhibited necroptosis in a dose-dependent way. The primary CD4+T cells were infected with HIV-1NL4-3 (5 ng HIV-1 p24 per 106 cells), with different concentrations of Nec-1 or NSA treatment. After 4 days, cells were harvested and analyzed by cell viability assay. Figure S4 - Detection the knockdown efficacy of target siRNAs. The primary CD4+T cells were first infected with HIV-1NL4-3 (5 ng HIV-1 p24 per 106 cells). After washing twice with PBS, infected cells were transfected with 30 nM siRNA and cultured in 24 well-plate. After 4 days, cells were harvested to extract RNA and the mNRA expression level of target genes were detected by RT-PCR. Figure S5 - The kinetics of viral infection is similar in both Jurkat cell lines. The wild-type and FADD-/-Jurkat cells were infected with HIV-1NL4-3 (5 ng HIV-1 p24 per 106 cells). After 4 and 7 days, cell supernatant was harvested and analyzed by p24 ELISA assay. *pYU2 (5 ng HIV-1 p24 per 106 cells) and then cultured in conditioned RPMI 1640 medium. After 4 or 7 days, the uninfected and infected cells were both harvested and analyzed by flow cytometry. B. Cell supernatant was harvested and analyzed by p24 ELISA assay. *p+T cells were infected with HIV-1NL4-3 (5 ng HIV-1 p24 per 106 cells). After 4 days, cell supernatant were collected and analyzed by TNF-α ELISA kit. *p (RAR)