t viride atcc 13233  (ATCC)

Journal: Frontiers in Immunology

Article Title: Cellular sensor DAP5 decodes Betacoronaviral NSP5 to drive virus-induced senescence

doi: 10.3389/fimmu.2026.1768183

Figure Lengend Snippet: Analysis of apoptosis and senescence in ACE2-HEK293T cells following SARS-CoV-2 infection. (A) Western blot analysis of caspase 3 cleavage in ACE2-HEK293T cells infected with SARS-CoV-2 (MOI = 0.1) for 24 hours. Caspase 3 activator raptinal (5 μM) was used as a positive control, with treatment for 6 hours. Viral infection was confirmed by Nucleocapsid (N) detection. The black asterisk indicates cleaved caspase 3. (B, C) Representative flow cytometry analysis (B) and statistical analysis (C) of apoptosis using Annexin V and PI staining in ACE2-HEK293T cells infected with SARS-CoV-2 (MOI = 0.1) at 0, 12, 24, and 48 hours post-infection. (D) The β-galactosidase activity of cells at 48 hours post-infection with SARS-CoV-2 (MOI = 0.1). Senescent cells are stained blue, as indicated by arrows. (E) The expression of senescence-associated genes CDKN1A , CDKN2A , IL1A , and IL8 in ACE2-HEK293T cells infected with SARS-CoV-2 (MOI = 0.1). (F) Western blot analysis of p21, p16, Lamin B1 and Nucleocapsid (N) expression in ACE2-HEK293T, ACE2-A549, Caco-2, and ACE2-MEF cells, either mock-treated or infected with SARS-CoV-2 at an MOI of 0.1 for 48 hours. (G) ELISA analysis of secreted IL-8 in cell culture supernatants of ACE2-A549 and Caco-2 cells at 48 hours after mock treatment or infection with SARS-CoV-2 (MOI = 0.1). Each experiment was independently repeated three times with similar results, and the representative images were shown. Mean ± SEM. ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: The hACE2 transgenic C57BL/6 mice were purchased from Cyagen and used to extract MEF cells (ACE2-MEF in the article).

Techniques: Infection, Western Blot, Positive Control, Flow Cytometry, Staining, Activity Assay, Expressing, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: bioRxiv

Article Title: Differential assembly of RNP granules via activation of distinct dsRNA sensors by adenovirus mutants

doi: 10.64898/2026.02.17.706363

Figure Lengend Snippet: ( A-D ) A549 RNase L KO cells treated with poly(I:C) (1 µg/mL for 6 h) or infected with ΔE4 (MOI 10 for 40 h) were stained for different granule markers. ( A ) UBAP2L (green) and FXR1 (magenta) show colocalization of both proteins to SGs induced by poly(I:C) and RLB-like granules induced during Δ E4 infection. Alternatively, the co-staining of G3BP1 with eIF4G1 ( B ) or Dcp1b ( C ) confirms Δ E4-induced RLBs do not colocalize with SG-unique proteins or P-bodies. ( D ) Staining for G3BP1 (green) and detection of polyadenylated RNA by FISH (magenta) shows recruitment of poly(A)+ RNA to both SGs and Δ E4-induced granules. ( E ) A549 parental and RNase L KO cells ( F ) were transfected with poly(I:C) (1 µg/mL for 6 h) or infected with Δ E4 (MOI 10 for 40 h). Cells were treated with DMSO or cycloheximide (CHX) 100 µg/mL to block translation and stress granule assembly. Fixed cells were stained for G3BP1 (green) and PABPC1 (magenta), with DAPI staining nuclei (blue). CHX treatment does not block poly(I:C) induced RLBs in parental cells, but blocks SG assembly in RNase L KO cells. Δ E4 infection induced RLB-like granules independently of CHX treatment in both cell lines. Scale bar = 15 µm (upper panels), 2 µm (cropped panels).

Article Snippet: The following primary antibodies were used for cellular proteins: Total PKR (Abcam ab32506, WB: 1:1,000), pT446 PKR (Abcam ab32036, WB: 1:1,000), total eIF2α (Cell Signaling 5324S, WB 1:1,000), pS51 eIF2α (Sigma-Aldrich SAB5701788, WB 1:500), GAPDH (GeneTex GTX100118, WB 1:10,000), IRF3 (Thermo Fisher 51-3200, WB 1:1,000), pS386 IRF (GeneTex GTX62132, 1:500), RNase L (Santa Cruz sc-74405, 1:500), G3BP1 mouse (DB 611126, IF 1:400), G3BP1 rabbit (Proteintech 13057-2-AP, IF 1:400), PABPC1 (Abcam ab21060, IF 1:600), UBAP2L (Bethyl Laboratories A300-534A-T, IF 1:100), FXR1 (EMD Millipore 05-1529, IF 1:100), eIF4G1 (Abcam ab2609, IF 1:100), FAM120A (Sigma-Aldrich HPA019734, IF 1:500), FMRP (EMD Millipore 05-1235, IF 1:250), Caprin1 (Proteintech 15112-1-AP, IF 1:200), Ago2 (Abnova H00027161-M01, IF 1:100), PRRC2C (Thermo Scientific PA5-55936, IF 1:100), LSm14A (Proteintech 18336-1-AP, IF 1:100), Dcp1b (Cell Signaling 13233S, IF 1:100).

Techniques: Infection, Staining, Transfection, Blocking Assay