human monocyte cell line thp 1 tib 202  (ATCC)

Journal: Cell Reports

Article Title: Nuclear import of malaria RNA rewires splicing in host immune cells

doi: 10.1016/j.celrep.2026.116953

Figure Lengend Snippet: Pf -EVs contain mRNAs that are associated with ribosomes in host monocytes (A) Concentrations of lipophilic dye (R18) and RNA cargo (SytoRNA) for Pf- EVs and uRBC-EVs ( n = 3, color coded, ∗ p < 0.05, two-tailed, unpaired t test); the dotted lines connect uRBC-EVs and Pf -EV samples from the same experiment. The data are also shown as fold changes ( Pf /uRBC EVs), with uRBC-EVs values normalized to an average of one. (B) Spectral signatures of unlabeled and labeled Pf -EVs acquired on spectral flow cytometer. (C) Illustration demonstrating the three enriched ETRAMPs encapsulated within Pf -EVs created with BioRender. (D) qPCR quantification of eleven ETRAMP mRNAs in Pf -EVs. Displayed are the Ct values; dots represent the six biological replicates. The dotted line indicates the threshold above which gene expression is considered undetectable (Ct = 37). (E) qPCR quantification of ETRAMP11.2 , ETRAMP2 , and ETRAMP14.1 in THP-1 cells treated with Pf -EVs for 1, 4, 16, and 24 h. Expression levels were normalized to the HPRT housekeeping gene and to the 1 h time point. Dots represent the three biological replicates. The dashed line indicates the baseline expression at 1 h, used as the reference for comparison across time points. (F) Polysome profiles of untreated THP-1 cells and THP-1 cells treated with Pf -EVs for 4 h. Shown are data from one experiment representative of three biological repeats. (G) qPCR analysis of the HPRT and ETRAMP11.2 mRNAs in the free (untranslated transcripts), light (transcripts translated at an intermediate level), and heavy (highly translated transcripts) polysomal fractions of THP-1 cells treated with Pf -EVs or untreated (UT) for 4 h ( n = 3), ∗ p < 0.05. The housekeeping gene HPRT served as a control. Error bars indicate SEM between the biological replicates.

Article Snippet: Human monocytic cell line THP-1 , ATCC , TIB-202.

Techniques: Two Tailed Test, Labeling, Flow Cytometry, Gene Expression, Expressing, Comparison, Control

Journal: Cell Reports

Article Title: Nuclear import of malaria RNA rewires splicing in host immune cells

doi: 10.1016/j.celrep.2026.116953

Figure Lengend Snippet: Pf mRNAs are rapidly internalized into host cell nuclei and are bound by endogenous RNA-binding proteins (A) Western blot analyses of nuclear and cytoplasmic fractions from Pf -EV-treated and untreated THP-1 cells for markers of the nucleus (lamin B2), cytoplasm (GAPDH), early endosomes (Rab 5), late endosomes (Rab 7), and endoplasmic reticulum (calnexin). (B) Representative qPCR analysis of nuclear and cytoplasmic fractions from Pf -EV-treated and untreated THP-1 cells at the indicated time points. Relative transcript levels were calculated using the standard curve method and normalized to the levels of U2 small nuclear RNA for the nuclear fraction, RPS11 for the cytoplasmic fraction, and to the 0.5 h time point. Shown is a qPCR analysis representative of three biological replicates. (C) Representative bright field (BF) and fluorescence confocal microscopy images of Pf -EV-treated and untreated THP-1 cells using HCR smFISH probes designed to identify ETRAMP11.2 (red) and ETRAMP2 (yellow). Hoechst was used to stain nuclei (blue), and carboxyfluorescein succinimidyl ester (CFSE) (green) was used to stain the cells. Scale bars represent 10 μm. (D) Quantification of the HCR smFISH probe signals (i.e., number of dots) using semiautomated Imaris software. Following segmentation of nuclei, cytoplasm, and FISH spots, the proportion of nuclear signal was calculated, with the cytoplasmic proportion defined as one minus the nucleus proportion. Statistical significance was assessed by comparing the nuclear proportions to an expected mean of 0.5 using a one-sample t test. Data represent the mean of three independent biological experiments; (ETRAMP11.2, p = 2.82 × 10 −7 and ETRAMP2, p = 6.26 × 10 −4 ); ∗∗∗ p < 0.001 and ∗∗∗∗ p < 0.0001. (E) Representative confocal microscopy images of Pf -EV-treated and untreated THP-1 cells using HCR smFISH probes designed to identify human β-actin mRNA, which served as positive host RNA control (green) and ETRAMP11.2 mRNA (red). Yellow arrows indicate ETRAMP11.2 in the nuclei of the monocyte. Hoechst was used to stain nuclei (blue). Scale bars represent 10 μm. (F) THP-1 cells were treated with Pf -EVs for 1 h. After UV crosslinking or not, biotinylated probes against ETRAMP11.2 mRNA were added to lysates, and the probes and associated mRNA and protein were precipitated with streptavidin-coated magnetic beads. Proteins were subjected to label-free quantitative mass spectrometry. Volcano plot of differentially expressed proteins precipitated in cells subjected to UV-mediated crosslinking and unirradiated cells. The results presented were significant based on a t test with permutation-based FDR calculation by log2 fold change (difference) and significance (−log p ) using the Perseus software. Protein quantification was based on four independent biological experiments.

Article Snippet: Human monocytic cell line THP-1 , ATCC , TIB-202.

Techniques: RNA Binding Assay, Western Blot, Fluorescence, Confocal Microscopy, Staining, Software, Control, Magnetic Beads, Mass Spectrometry

Journal: Cell Reports

Article Title: Nuclear import of malaria RNA rewires splicing in host immune cells

doi: 10.1016/j.celrep.2026.116953

Figure Lengend Snippet: Pf -EV treatment changes the proteomic profile of monocytes (A) Plot of differential protein expression in THP-1 cells treated with Pf -EVs for 16 h versus control untreated cells (UT). The x axis is the ratio between the amount of a protein detected in the existing (“light” isotope) pool and that in the newly synthesized (“heavy” isotope) pool (H/L) of the Pf -EV-treated samples and the H/L of the untreated sample (log2). The y axis represents the fold change between the total intensities of the two groups (log2). Data are the averages of the three repeats, and missing values were replaced by the minimal intensity in the experiment. In the upper right quadrant, the newly synthesized proteins elevated post Pf -EV treatment that were also elevated in the total intensity are marked in red. In the lower left quadrant, the newly synthesized proteins decreased post Pf -EV treatment, which were also decreased in the total intensity, are marked in blue. (B) Network interactions of proteins downregulated in THP-1 cells upon Pf -EV treatment as compared to untreated cells by search tool for the retrieval of interacting genes/proteins (STRING) analysis. (C) Gene Ontology (GO) annotation enrichments of human downregulated proteins in THP-1 cells upon Pf -EV treatment as compared to untreated cells. Count refers to the number of proteins that were annotated with a particular term. (D) Quantification of indicated mRNA levels in THP-1 cells treated with Pf -EVs for 1, 2, 3, 4, and 16 h. HPRT served as a control. Data are presented as the fold induction over untreated control (UT – 1 h) and are means and standard errors of the means of three biological replicates. ΔCT values per gene were compared with a linear model (two-way ANOVA) testing the effect of treatment, time, treatment and time interaction, and batch ( RHOT2 , p = 0.0049; TEX264 , p = 0.0460; SCLY , p = 0.0176; and FN3KRP , p = 0.0411), ∗ p < 0.05 and ∗∗ p < 0.01.

Article Snippet: Human monocytic cell line THP-1 , ATCC , TIB-202.

Techniques: Expressing, Control, Synthesized

Journal: Cell Reports

Article Title: Nuclear import of malaria RNA rewires splicing in host immune cells

doi: 10.1016/j.celrep.2026.116953

Figure Lengend Snippet: Splicing is altered in monocytes after treatment with Pf -EVs as revealed by RNA-seq analysis (A) Volcano plot of all AS events between Pf -EV-treated and untreated THP-1 cells. Shown are significant results based on Δ percent spliced in (PSI) (inclusion level difference), and significance (−log p ) determined using rMATS-turbo. (B) Volcano plots of individual AS events (skipped exons [SE], retained introns [RI], alternative 3′ splice sites [A3SS], alternative 5′ splice sites [A5SS], and mutually exclusive exons [MXE]) between Pf -EV-treated and untreated THP-1 cells. Shown are significant results based on ΔPSI (inclusion level difference), and significance (−log p ). (C) Raw counts of each AS event. (D) KEGG pathways enrichment analysis of AS events from rMATS between Pf -EV-treated and untreated THP-1 cells. (E) Detailed sashimi plots depicting the exon skipping events of ENOPH1 and KEAP1 transcripts. ΔPSI was calculated as the ratio between the PSI of Pf -EV-treated cells and that of untreated THP-1 cells. (F) Volcano plot of differentially expressed genes between Pf -EV-treated and untreated THP-1 cells. Shown are significant results based on log2 fold change (difference), and significance (−log p ) determined using DEseq2 (version 1.38.3). Quantification was based on two independent biological experiments. (G) Network interactions of upregulated genes in THP-1 cells upon Pf -EV treatment as compared to untreated cells by STRING analysis. (H) qPCR analysis of the upregulated genes in Pf -EV-treated versus untreated THP-1 cells. HPRT served as a housekeeping control gene. Error bars indicate SEM between the three biological replicates. ΔCT values per gene were compared with a t test ( CCL4 , p = 0.009; CXCL10 , p = 0.022; ICAM-1 , p = 0.009; CCL3 , p = 0.056; and IL-1β , p = 0.007); ∗ p < 0.05 and ∗∗ p < 0.01.

Article Snippet: Human monocytic cell line THP-1 , ATCC , TIB-202.

Techniques: RNA Sequencing, Control

Journal: Cell Reports

Article Title: Nuclear import of malaria RNA rewires splicing in host immune cells

doi: 10.1016/j.celrep.2026.116953

Figure Lengend Snippet: Pf -EVs induce a pro-inflammatory response in monocytes (A) Network interactions of proteins upregulated in THP-1 cells upon Pf -EV treatment as compared to untreated cells by STRING analysis. (B) GO annotation enrichments of human upregulated proteins in THP-1 cells upon Pf -EV treatment as compared to untreated cells. Count refers to the number of proteins that were annotated with a particular term. (C) Relative expression of CD80 , TNF-α , IL-6 , and IL-12 mRNAs in MDMs. Monocytes isolated from healthy donors were treated with M-CSF only; with M-CSF plus Pf -EVs; or with M-CSF, IL-4, and IL-10. Cells were harvested, and RNA was extracted and subjected to qPCR analysis. GAPDH served as a housekeeping control gene. Data are presented as the fold induction over untreated control (M-CSF – 6 h) and represent the mean and standard error of the mean of three biological replicates. ΔCt values were compared using an ANOVA model, accounting for treatment, time, and batch ( CD80 : Pf -EVs p = 0.0003 and IFN-γ [both] p < 0.0001; IL-6 : Pf -EVs p = 0.0002 and IFN-γ [both] p < 0.0001; IL-12 : Pf -EVs p = 0.0223 and IFN-γ [both] p < 0.0001; and TNF-α : Pf -EVs p = 0.0562, IFN-γ [6 h] p < 0.0001, and IFN-γ [24 h] p = 0.0004); ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. (D) Model of Pf mRNA nuclear localization and binding by host cell proteins ACIN1 and PNN, leading to splicing alterations, endogenous protein downregulation, and upregulation of the pro-inflammatory response in host monocytes. Illustration created with BioRender.

Article Snippet: Human monocytic cell line THP-1 , ATCC , TIB-202.

Techniques: Expressing, Isolation, Control, Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Improving Growth Dynamics of Faecalibacterium prausnitzii by Exposure to Prebiotics

doi: 10.3390/ijms27041698

Figure Lengend Snippet: TNF expression in THP-1 cells stimulated with F. prausnitzii culture supernatants derived from different prebiotics. The gene expression level of TNF-α was normalized with GAPDH as a control housekeeping gene and calculated using the 2 −ΔΔCt method. Values of triplicate experiments are demonstrated as mean ± SE. Statistical differences among groups were determined using one-way ANOVA, with p < 0.05 considered significant.

Article Snippet: To determine whether the supernatant of F. prausnitzii following culture with various prebiotics could modulate immune cell response, THP-1 (human monocytic cell line) cells were obtained from ATCC (American Type Culture Collection: TIB-202) and grown in suspension in RPMI + Glutamax supplemented with 10% ( v / v ) FBS in a humidified 37 °C, 5% CO 2 incubator.

Techniques: Expressing, Derivative Assay, Gene Expression, Control