targeting control shrna  (Addgene inc)

Journal: iScience

Article Title: Translation factor eIF4G2 directs CD8 + T cell lineage commitment by selectively enabling the IL-7 receptor response

doi: 10.1016/j.isci.2026.115313

Figure Lengend Snippet: eIF4G2 post-transcriptionally sustains γc expression via its mRNA UTRs (A–C) Analysis in primary CD4 + CD8 lo transitional thymocytes. (A) Western blot analysis of γc and IL-7Rα protein levels. (B and C) Quantitative RT-PCR analysis of Il2rg (B) and Il7r (C) mRNA levels ( n = 3, ns p > 0.05, ∗ p < 0.05). (D–H) Mechanistic dissection in 293T cells. (D) Western blot of γc protein in control and EIF4G2 knockdown 293T cells transfected with an IL2RG coding sequence construct containing its native 5′ and 3′ UTRs. (E) Corresponding IL2RG mRNA levels measured by RT-qPCR ( n = 3, ns p > 0.05) . (F and G) Assessment of γc protein stability ( n = 3, ns p > 0.05). (F) Representative western blots and (G) quantification of γc protein levels over time following cycloheximide (CHX) treatment in si-control and si- EIF4G2 293T cells ( n = 3, ns p > 0.05). (H) Western blot of γc protein in si-control and si- EIF4G2 293T cells transfected with an IL2RG CDS construct lacking UTRs. Data are representative of at least two independent experiments. Bar graphs show mean ± SEM and unpaired Students’ t test was used to perform the statistical analysis.

Article Snippet: Cells were transfected either with a non-targeting control siRNA (si-Control) or human EIF4G2 targeting siRNA (si- EIF4G2 ) (Sangon Biotech) using Lipofectamine RNAiMAX transfection reagent (Invitrogen).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Dissection, Control, Knockdown, Transfection, Sequencing, Construct

Journal: Poultry Science

Article Title: eIF3m promotes fowl adenovirus serotype 4 replication via interacting with ORF1B protein

doi: 10.1016/j.psj.2026.106566

Figure Lengend Snippet: Knockdown of eIF3m suppressed FAdV-4 replication in vitro. (A and B) LMH cells transfected with siRNA targeting at eIF3m or siNC as a negative control were collected at 24 h for qRT-PCR (A) and at 48 h for western blot (B) to evaluate the knockdown effect. (C and D) LMH cells transfected with 100nM/well siRNA or siNC were infected with CH/HNJZ/2015 at an MOI of 0.01. Western blot (C) and virus titration (D) were analyzed to investigate virus growth kinetics. The data shown represent the means±SD, and the experiments were repeated three times. The statistics analysis was performed by 2-way ANOVA. *, P < 0.05, **, P < 0.01.

Article Snippet: Based on the chicken eIF3m sequence obtained from GenBank, three siRNAs along with a non-targeting control (siNC) were synthesized by Shanghai Sangon Biotech.

Techniques: Knockdown, In Vitro, Transfection, Negative Control, Quantitative RT-PCR, Western Blot, Infection, Virus, Titration