Journal: International Journal of Molecular Sciences

Article Title: Unfolding Immune Dysregulation in COPD: Identification of a Three-Gene Signature and Functional Validation of TCF7 in Human Lung Tissue and T Lymphocytes

doi: 10.3390/ijms27104231

Figure Lengend Snippet: TCF7 regulates pro−caspase−8 expression in T lymphocytes and is significantly reduced in COPD. ( A ) Immunofluorescence co−staining of control human lung tissue displaying separate channels for DAPI (blue), caspase−8 (green), TCF7 (red), and the merged image. Scale bar is 50 μm. ( B ) Immunofluorescence co−staining of COPD human lung tissue displaying separate channels for DAPI (blue), caspase−8 (green), TCF7 (red), and the merged image. Note the marked reduction in both TCF7 and caspase−8 signals compared to the control. Scale bar is 50 μm. ( C ) Representative Western blot images of TCF7 (50 kDa), pro−caspase−8 (55 kDa), and internal control β−tubulin (55 kDa) in wild type (WT) and TCF7 knockout (KO) Jurkat T cells. ( D ) Quantitative densitometric analysis of TCF7 protein levels comparing WT and KO groups. ( E ) Quantitative densitometric analysis of pro−caspase−8 protein levels comparing WT and KO groups. ( F ) Representative Western blot images of TCF7 and β−tubulin in primary T lymphocytes isolated from the peripheral blood of healthy donors (Control) and patients with COPD (Model). ( G ) Quantitative densitometric analysis of TCF7 protein levels in human primary T lymphocytes. ( H ) Representative Western blot images of TCF7 and β−tubulin protein levels in Jurkat T cells across four experimental conditions including Control, shRNA, shRNA plus TCF7 Rescue construct, and shRNA plus Empty Vector. ( I ) Quantitative densitometric analysis of TCF7 protein levels across the four experimental rescue groups. ( J ) Representative Western blot images of pro−caspase−8 and β−tubulin protein levels across the same four experimental conditions in Jurkat T cells. ( K ) Quantitative densitometric analysis of pro−caspase−8 protein levels across the four experimental rescue groups. Data in the bar charts are presented as mean ± SD ( n = 4 for primary human cells, n = 3 for cell line experiments). Statistical significance was assessed using Student’s t test with Welch’s correction where appropriate (* p < 0.05, *** p < 0.001, ns indicates not significant).

Article Snippet: Short hairpin RNA targeting human TCF7 (shRNA) and a scramble control shRNA were purchased from OriGene with Cat.No.TR30004.

Techniques: Expressing, Immunofluorescence, Staining, Control, Western Blot, Knock-Out, Isolation, shRNA, Construct, Plasmid Preparation

Journal: bioRxiv

Article Title: Deep Learning-Driven Discovery of Mitochondrial Factors Modulating Influenza A Virus Infection

doi: 10.64898/2026.02.25.707858

Figure Lengend Snippet: a, IFN-βmRNA relative expression. HeLa cells were transfected with siRNAs targeting individual mitochondrial genes or a non-targeting control (siControl), followed by infection with influenza A virus. IFN-β mRNA levels were measured by quantitative PCR (qPCR), using PUM1 as the normalization reference gene. b, Influenza virus PA and HA vRNAs relative expression. HeLa cells were transfected and infected as described above. vRNA levels were assessed by qPCR and normalized to PUM1 mRNA expression. c, Influenza virus production. Viral titers in the supernatant were quantified by plaque assay following infection of siRNA-transfected cells. All data represent the mean ± SEM (a, b) or ± s.d. (c) of three biological replicates. Genes with a fold change > 2 are indicated by patterned bars. Statistical analysis was performed using one-way ANOVA, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The cells were collected after 48 hours to analyse for knockdown efficiency or used for further experiments. siRNA used: Hsp60 (HSPD1) Human siRNA Oligo Duplex (OriGene, locus ID 3329) AccuTarget Predesigned Human ETHE1 siRNA (Bioneer, locus ID 23474) AccuTarget Predesigned Human GRP75 siRNA (Bioneer, locus ID 3313) LETM1 Human siRNA Oligo Duplex (OriGene, locus ID 3954) AccuTarget Predesigned Human LONP1 siRNA (Bioneer, locus ID 9361) OXA1L Human siRNA Oligo Duplex (OriGene, locus ID 5018) AccuTarget Predesigned Human MPPB siRNA (Bioneer, locus ID 9512) AccuTarget Predesigned Human TIM44 siRNA (Bioneer, locus ID 10469) AccuTarget Predesigned Human SQOR siRNA (Bioneer, locus ID 58472) Non-targeting scramble siRNA (siControl) was purchased from OriGene.

Techniques: Expressing, Transfection, Control, Infection, Virus, Real-time Polymerase Chain Reaction, Plaque Assay