Journal: Redox Report : Communications in Free Radical Research

Article Title: Elucidating the protective role of quercetin against lipopolysaccharide-induced necroptosis in broiler thymus: insights from Nrf2/PERK signaling based on network pharmacology and experimental validation

doi: 10.1080/13510002.2026.2663621

Figure Lengend Snippet: QUE attenuated necroptosis through Nrf2/PERK under LPS exposure. (A and B) IF results for RIPK3 and MLKL in thymus tissues ( n = 20 broilers per group). (C and D) The detection for mRNA and protein levels of necroptosis factors (RIPK1, RIPK3, Caspase8, MLKL, and P -MLKL) in thymus tissues ( n = 20 broilers per group). (E and F) IF results for necroptosis marker factors (RIPK3 and MLKL) in MSB-1 cells and quantitative analysis ( n = 3 independent cell culture experiments). (G and I) The transcription and protein levels of necroptosis factors, including RIPK1, RIPK3, Caspase8, MLKL, and P -MLKL, in MSB-1 cells ( n = 3 independent cell culture experiments). All the data are displayed as the means ± SD ( n = 3). The statistically significant differences are not denoted with the same letters, such as ‘a, b, c, and d’.

Article Snippet: Following dewaxing, the slices were subjected to antigen retrieval and blocked with 3% BSA at room temperature for 30 min. Then, the slices were incubated with PERK (GB11507-100, 1:200, Rabbit pAb, Servicebio, Wuhan), MLKL (GB155699-100, 1:5000, Rabbit pAb, Servicebio, Wuhan), and RIPK3 (bs-3551R, 1:100, Rabbit pAb, Bioss, Beijing) primary antibodies, secondary antibodies, and stained with DAPI solution.

Techniques: Marker, Cell Culture

Journal: NAR Molecular Medicine

Article Title: Human pneumovirus induces IFN-dependent expression of the immune-responsive gene 1 and is inhibited by 4-octyl itaconate in human macrophages

doi: 10.1093/narmme/ugag017

Figure Lengend Snippet: TBK1 and NF-κB positively regulate IRG1 levels in HMPV-infected human macrophages. MDMs were incubated with 5 or 10 μM of the TBK1 inhibitor BX795 for 30 min before infection with HMPV for 24 h. ( A ) IRG1 and IFN-β mRNA levels ( n ≥ 3) were analysed by qRT-PCR (left panels), while in panel ( B ) IRG1 and GAPDH protein levels were determined by immunoblotting, quantified, and presented with SD relative to uninfected cells treated with DMSO ( n = 2; right panel). (C–K) MDMs were transfected with siRNAs targeting the NF-κB subunit RELA (C–E), IRF1 (F–H), RIPK3 (I–K), or control siRNA (siNTC) before infection with HMPV for 6, 9, or 24 h and analysis of IRG1 mRNA levels by qRT-PCR or IRG1, RelA/p65, IRF1, RIPK3, or GAPDH protein levels by immunoblotting. IRG1 mRNA levels were assessed relative to siNTC-transfected, uninfected MDMs for all experiments. ( C, E ) IRG1 mRNA after 6 h ( n = 3) or 24 h HMPV ( n = 3). ( D ) Protein levels of IRG1, RelA/p65, and GAPDH ( n = 3) after 9 h HMPV. ( F, H ) IRG1 mRNA after 6 h ( n = 4) or 24 h of infection ( n = 3). ( G ) Protein levels of IRG1, IRF1, and GAPDH ( n = 3) after 9 h HMPV. ( I, K ) IRG1 mRNA after 6 h ( n = 3) or 24 h of infection ( n = 3). ( J ) Protein levels of IRG1, IRF1, and GAPDH ( n = 3) after 9 h HMPV. ( L ) MDMs were incubated with 100 nM of the RIPK3 inhibitor GSK 872 for 1 h before infection with HMPV for 24 h. IRG1 mRNA levels were normalized relative to uninfected cells treated with DMSO ( n = 2). Single comparison between control (NTC) siRNA and target-siRNA conditions were calculated using paired t -test with Tukey post-hoc test. * P < .05, ** P < .01, *** P < .001, **** P < .0001; ns = non-significant. See also

Article Snippet: The primary human antibodies IRG1, IRF1, RIPK3, NF-κB p65, phospho-STAT1 (Tyr701), STAT1, and Nrf2 were purchased from Cell Signalling Technology.

Techniques: Infection, Incubation, Quantitative RT-PCR, Western Blot, Transfection, Control, Comparison