Journal: bioRxiv

Article Title: The zinc metalloprotease ZMPSTE24 binds a distinct topological isoform of the tail-anchored protein IFITM3

doi: 10.64898/2026.02.27.708584

Figure Lengend Snippet: (A) Assessing loss of catalytic activity for ZMPSTE24 HExxH…E mutants. Flag-tagged wild-type and mutant ZMPSTE24 plasmids were co-transfected with a myc-tagged prelamin A plasmid (myc-LMNA (431-664) ) in ZMPSTE24 knockout HEK293 cells to evaluate ZMPSTE24 proteolytic activity. Flag-ZMPSTE24, prelamin A and its cleaved product, mature lamin A, were detected by Western blotting and are indicated. All active site mutants block cleavage of the prelamin A construct. (B) Co-IP’s of endogenous IFITMs by ZMPSTE24 and ZMPSTE24 E336A . HEK293 cells transfected with vector, Flag-ZMPSTE24 or Flag-ZMPSTE24 E336A were induced with 150 units of interferon β for 18 hours prior to immunoprecipitation with anti-Flag agarose. Whole cell lysate (WCL) and IP’d proteins were resolved by SDS-PAGE analyzed by Western blotting with anti-Flag (ZMPSTE24), anti-IFITM1, IFITM2 or IFITM3 antibodies. (C) Comparing co-IPs of IFITMs by Flag-ZMPSTE24 E336A and Flag-IFITM3. Cells transfected with vector, Flag-ZMPSTE24, Flag-ZMPSTE24 E336A or Flag-IFITM3 and the indicated myc-tagged IFITM proteins were subjected to co-IP as described. Positions of ZMPSTE24 and IFITM proteins are indicated. Lanes 9-12 were used to make . (D) Flag-tagged IFITM3 co-IPs myc-ZMPSTE24 E336A more efficiently than wild-type myc-ZMPSTE24. HEK293 cells were transfected with Flag-IFITM3 and myc-ZMPSTE24 or myc-ZMPSTE24 E336A . Proteins were co-IP’d with anti-Flag agarose prior to SDS-PAGE and Western blotting. Positions of Flag-IFITM3 and myc-ZMPSTE24 proteins are indicated.

Article Snippet: Anti-IFITM1 (Proteintech, 11727-3-AP) was used at 1:1000.

Techniques: Activity Assay, Mutagenesis, Transfection, Plasmid Preparation, Knock-Out, Western Blot, Blocking Assay, Construct, Immunoprecipitation, SDS Page, Co-Immunoprecipitation Assay

Journal: bioRxiv

Article Title: Interferon Beta Drives Therapy Resistance in a Patient-Derived Model of High-Grade Serous Ovarian Cancer

doi: 10.64898/2026.01.27.699126

Figure Lengend Snippet: A. RT-qPCR analysis of IFN-1 RNA levels. B . Secreted IFN-1 protein levels detected in culture supernatant via HEK-Blue SEAP reporter assay. C . RT-qPCR analysis of IRDS genes. Dots indicate genes known to be transcribed by u-ISGF3. D . Intracellular flow cytometry of IFITM1, total STAT1, and pSTAT1(y701). One representative replicate shown. Results are displayed as n=3, unless otherwise noted, and presented as the means ± SD. Statistical significance was determined using the unpaired t-test. p-values: p ≤ 0.05 (*).

Article Snippet: Cells were also stained using a two-step approach first staining with IFITM1 (99969S; Cell Signaling Technology) diluted in 0.5% BSA for 1 hour at room temperature before rinsing and staining with AlexaFluorTM 647 goat anti-rabbit IgG (H+L) secondary antibody (A21245; Fisher Scientific) at room temperature for 30 minutes, washed, and resuspended in FACS stain.

Techniques: Quantitative RT-PCR, Reporter Assay, Flow Cytometry

Journal: bioRxiv

Article Title: Interferon Beta Drives Therapy Resistance in a Patient-Derived Model of High-Grade Serous Ovarian Cancer

doi: 10.64898/2026.01.27.699126

Figure Lengend Snippet: A. The IC50 of SE and CR cells was significantly higher following IFNβ treatment. B . Morphology change was determined by measuring the aspect ratio (width/legth) of 60 cells from each sample (20 cells per photo, 2 photos per well, 3 wells). Scale bar = 50µm. C . BrdU staining revealed cell cycle arrest in G0-G1 following IFNβ treatment, as measured by flow cytometry. Data represent one representative replicate. D . Proliferation was detected by counting individual wells of parental and IFNβ treated cells over time. E . RT-qPCR was used to analyze STAT1, IFITM1, and PLSCR1 following 6 passages and 12 weeks following consistent low-level IFNβ treatment. F . Intracellular flow cytometry was used to measure STAT1 and pSTAT1 protein levels following chronic IFNβ treatment. One representative replicate shown. Results are displayed as n=3, unless otherwise noted, and presented as the means ± SD. Statistical significance was determined using the unpaired t-test. p-values: p ≤ 0.05 (*) and p ≤ 0.01 (**).

Article Snippet: Cells were also stained using a two-step approach first staining with IFITM1 (99969S; Cell Signaling Technology) diluted in 0.5% BSA for 1 hour at room temperature before rinsing and staining with AlexaFluorTM 647 goat anti-rabbit IgG (H+L) secondary antibody (A21245; Fisher Scientific) at room temperature for 30 minutes, washed, and resuspended in FACS stain.

Techniques: BrdU Staining, Flow Cytometry, Quantitative RT-PCR

Journal: Biomolecules and Biomedicine

Article Title: S-palmitoylation-related genes in Crohn’s disease: Bioinformatic identification and validation

doi: 10.17305/bb.2025.13221

Figure Lengend Snippet: Multi-cohort validation of ZDHHC23 and IFITM1 as diagnostic biomarkers for CD. (A) Significant downregulation of ZDHHC23 expression across multiple CD cohorts. (B) Significant upregulation of IFITM1 expression across multiple CD cohorts. (C) ROC curve illustrating the diagnostic utility of ZDHHC23 in CD detection. (D) ROC curve illustrating the diagnostic utility of IFITM1 in CD detection. Asterisks indicate statistical significance (*** P < 0.001; **** P < 0.0001). Abbreviations: CD: Crohn’s disease; ROC: Receiver operating characteristic.

Article Snippet: Membranes were blocked with 5% bovine serum albumin (BSA) for 1 h at room temperature and subsequently incubated overnight at 4 ∘ C with primary antibodies diluted as follows: GAPDH (1:4000, Proteintech), ZDHHC23 (1:1000, Epigentek), and IFITM1 (1:1000, Proteintech).

Techniques: Biomarker Discovery, Diagnostic Assay, Expressing

Journal: Biomolecules and Biomedicine

Article Title: S-palmitoylation-related genes in Crohn’s disease: Bioinformatic identification and validation

doi: 10.17305/bb.2025.13221

Figure Lengend Snippet: Visualization of immune cell infiltration. (A and B) The correlation matrix of infiltrating immune cell composition and proportions was generated using the CIBERSORT algorithm. (C and D) The correlation matrix of infiltrating immune cell composition and proportions was produced using the MCPcounter algorithm. (E and F) The correlation matrix of infiltrating immune cell composition and proportions was established using the QuanTIseq algorithm. (G) Correlation between ZDHHC23 expression and infiltrating immune cells, specifically M0 Macrophages, activated dendritic cells, and regulatory T cells. (H) Correlation between IFITM1 expression and infiltrating immune cells, including follicular helper T cells, regulatory T cells, and resting NK cells. Asterisks (*) indicate correlations that remained statistically significant after FDR correction for multiple testing across all gene–cell pairs within each algorithm ( q < 0.05). Abbreviations: CIBERSORT: Cell-type identification by estimating relative subsets of RNA transcripts; FDR: False discovery rate; NK: Natural killer.

Article Snippet: Membranes were blocked with 5% bovine serum albumin (BSA) for 1 h at room temperature and subsequently incubated overnight at 4 ∘ C with primary antibodies diluted as follows: GAPDH (1:4000, Proteintech), ZDHHC23 (1:1000, Epigentek), and IFITM1 (1:1000, Proteintech).

Techniques: Generated, Produced, Expressing

Journal: Biomolecules and Biomedicine

Article Title: S-palmitoylation-related genes in Crohn’s disease: Bioinformatic identification and validation

doi: 10.17305/bb.2025.13221

Figure Lengend Snippet: Experimental validation of ZDHHC23 and IFITM1 expression in LPS-treated HT-29 cells. The expression levels of ZDHHC23 and IFITM1 in HT-29 cells were validated through qRT-PCR analysis, which confirmed mRNA levels for ZDHHC23 (A) and IFITM1 (B). Western blot analysis (C) demonstrated protein expression differences between control and LPS-treated HT-29 cells ( n ═ 3 biological replicates). Densitometric quantification of protein expression was performed for ZDHHC23 (D) and IFITM1 (E). Asterisks indicate statistical significance (* P < 0.05; ** P < 0.01; *** P < 0.001). Abbreviations: LPS: Lipopolysaccharide; qRT-PCR: Quantitative reverse transcription PCR.

Article Snippet: Membranes were blocked with 5% bovine serum albumin (BSA) for 1 h at room temperature and subsequently incubated overnight at 4 ∘ C with primary antibodies diluted as follows: GAPDH (1:4000, Proteintech), ZDHHC23 (1:1000, Epigentek), and IFITM1 (1:1000, Proteintech).

Techniques: Biomarker Discovery, Expressing, Quantitative RT-PCR, Western Blot, Control, Reverse Transcription