human thp  (ATCC)

Journal: Materials Today Bio

Article Title: Targeting FN1 to overcome gemcitabine resistance in gallbladder cancer: Mechanistic insights and an iRGD-modified PEG-PLGA nanoparticle delivery strategy

doi: 10.1016/j.mtbio.2026.102877

Figure Lengend Snippet: Effect of FN1 knockdown on the immunosuppressive microenvironment in GBC-SD/GEM cells. Note: (A) Schematic of immunosuppressive cells and cytokines detection in tumor tissues; (B) FCM analysis of Tregs infiltration levels in GBC-SD/GEM cell xenograft tissues in various mouse groups; (C) FCM analysis of M2 and M1 macrophage infiltration levels in GBC-SD/GEM cell xenograft tissues from different mouse groups; (D-E) RT-qPCR analysis of immunosuppressive factors expression in GBC-SD/GEM cell xenograft tissues from various mouse groups; (F) Schematic of in vitro co-culture of GEM-resistant GBC cells with THP-1 and CD4 + T cells; (G) FCM analysis of Tregs levels in CD4 + T cells after co-culture with GBC-SD/GEM cells; (H) FCM analysis of M2 and M1 macrophage levels in THP-1 cells post co-culture with GBC-SD/GEM cells; (I) RT-qPCR analysis of IL-10 or CSF-1 expression in CD4 + T or THP-1 cells after co-culture with GBC-SD/GEM cells. In B-E, ∗ indicates p < 0.05 compared to the sh-NC + GEM group, each group consisting of 6 mice; in G-I, ∗ indicates p < 0.05 compared to the sh-NC group, # indicates p < 0.05 compared to the oe-NC group, experiments repeated three times.

Article Snippet: Human GBC cell lines NOZ (CC-Y1668) and GBC-SD (CC-Y1162) were obtained from Shanghai EK-Bioscience Biotechnology Co., Ltd. Human monocytic cells THP-1 (TIB-202) and 293T cells (CRL-3216) were purchased from the ATCC.

Techniques: Knockdown, Quantitative RT-PCR, Expressing, In Vitro, Co-Culture Assay

Journal: Materials Today Bio

Article Title: Targeting FN1 to overcome gemcitabine resistance in gallbladder cancer: Mechanistic insights and an iRGD-modified PEG-PLGA nanoparticle delivery strategy

doi: 10.1016/j.mtbio.2026.102877

Figure Lengend Snippet: Impact of NPs delivering si-FN1 on drug resistance and immune cell infiltration in GBC-SD/GEM cells. Note: (A) Schematic of the experimental setup for studying the impact of NPs delivering si-FN1 on GBC GEM resistance; (B-C) RT-qPCR (B) and Western Blot (C) analysis of FN1 and PI3K pathway protein expression in GBC-SD/GEM cells treated with NPs (si-FN1) and iRGD-NPs (si-FN1); (D) CCK-8 assay assessing the viability changes in GBC-SD/GEM cells after treatment with NPs (si-FN1) and iRGD-NPs (si-FN1); (E) Clonogenic assay evaluating colony formation in various groups of GBC-SD/GEM and NOZ/GEM cells; (F) FCM analysis of apoptosis in GBC-SD/GEM cells across different groups; (G) FCM analysis of Tregs levels in CD4 + T cells after co-culture with GBC-SD/GEM cells; (H) FCM analysis of M2 and M1 macrophage levels in THP-1 cells after co-culture with GBC-SD/GEM cells; (I) RT-qPCR analysis of IL-10 or CSF-1 expression in CD4 + T or THP-1 cells co-cultured with GBC-SD/GEM cells. ∗ indicates p < 0.05 compared to the NPs (si-NC) group, # indicates p < 0.05 compared to the NPs (si-FN1) group, experiments repeated three times.

Article Snippet: Human GBC cell lines NOZ (CC-Y1668) and GBC-SD (CC-Y1162) were obtained from Shanghai EK-Bioscience Biotechnology Co., Ltd. Human monocytic cells THP-1 (TIB-202) and 293T cells (CRL-3216) were purchased from the ATCC.

Techniques: Quantitative RT-PCR, Western Blot, Expressing, CCK-8 Assay, Clonogenic Assay, Co-Culture Assay, Cell Culture

Journal: iScience

Article Title: MALAT1 regulates human macrophage metabolism by interacting with HADHB

doi: 10.1016/j.isci.2026.115107

Figure Lengend Snippet: MALAT1 binds to HADHB (A) Schematic representation of the RNA immunoprecipitation (RIP) of LPS-stimulated THP-1-derived macrophages differentiated with PMA for 24 h. (B) Western blot of RIP samples using an anti -HADHB antibody, with IgG as a negative control. The experiment was repeated; n = 3 independent experiments (independent cell cultures on different days). (C) Top ten candidate non-coding RNAs binding to HADHB, ranked by peak score from RIP-seq analysis. The experiment was repeated; n = 3 independent experiments (independent cell cultures on different days). (D) Fold enrichment of MALAT1 in RIP samples. The experiment was repeated; n = 3 independent experiments (independent cell cultures on different days); data are presented as mean ± SD and analyzed with the two-tailed unpaired Student’s t test.

Article Snippet: Human THP-1 cell lines and Raw 264.7 cell lines were purchased from the American Type Culture Collection (ATCC, TIB-202, TIB-71).

Techniques: RNA Immunoprecipitation, Derivative Assay, Western Blot, Negative Control, Binding Assay, Two Tailed Test

Journal: iScience

Article Title: MALAT1 regulates human macrophage metabolism by interacting with HADHB

doi: 10.1016/j.isci.2026.115107

Figure Lengend Snippet: MALAT1 translocates to mitochondria via the HuR-MTCH2 axis and enhances HADHB thiolase activity (A) MALAT1 transcript levels in the nucleus, cytoplasm, and mitochondria, normalized to U6 (nuclear RNA), GAPDH (cytoplasmic RNA), and COX2 (mitochondrial RNA). The experiment was repeated; n = 9 independent experiments (independent cell cultures on different days); data are shown as mean ± SD and analyzed with the two-way ANOVA followed by Tukey’s post hoc test. (B–D) MALAT1 transcript levels in the nucleus, cytoplasm, and mitochondria of PMA-differentiated THP-1 macrophages stimulated with LPS for various durations. For each LPS stimulation time point, n = 9 independent experiments (independent cell cultures and stimulated on different days); data are shown as mean ± SD and analyzed with the one-way ANOVA followed by Tukey’s post hoc test. (E) Western blot of RIP samples from THP-1-derived macrophages stimulated with LPS for 24 h or 48 h, probed with anti -HuR and anti -MTCH2, input and IgG controls shown. For each LPS stimulation time point, n = 3 independent experiments (independent cell cultures and stimulated on different days). (F) Fold-change enrichment of MALAT1 in HuR and MTCH2 RIP samples from THP-1-derived macrophages after LPS stimulation (24 h and 48 h). The experiment was repeated; n = 3 independent experiments (independent cell cultures on different days); data are presented as mean ± SD and analyzed with the two-tailed unpaired Student’s t test. (G) Reciprocal RIP-western confirms HuR-MTCH2 association in THP-1-derived macrophages after 24- and 48-h LPS stimulation. For each LPS stimulation time point, n = 3 independent experiments (independent cell cultures and stimulated on different days). (H) MALAT1 transcript levels in siRNA-transfected THP-1-derived macrophages (the experiment was repeated; n = 9 independent experiments) and MALAT1 overexpression THP-1 cells (the experiment was repeated n = 3 independent experiments), normalized to GAPDH and compared with siNC. Data are shown as mean ± SD and analyzed with one-way ANOVA followed by Dunnett’s post hoc test. (I) Schematic representation of FAO. HADHB, the β-subunit of the mitochondrial trifunctional protein (MTP), possesses thiolase activity and catalyzes the final step of FAO. Diagram created with BioRender. (J) Flow cytometric analysis of HADHB thiolase activity measured by mean fluorescence intensity (MFI) in THP-1-derived macrophages transfected with siRNA (siMALAT1) or stably overexpressing MALAT1 (Oe- MALAT1 ). The experiment was repeated; n = 3 independent experiments (independent cell cultures on different days); data are shown as mean ± SD and analyzed with the two-tailed unpaired Student’s t test.

Article Snippet: Human THP-1 cell lines and Raw 264.7 cell lines were purchased from the American Type Culture Collection (ATCC, TIB-202, TIB-71).

Techniques: Activity Assay, Western Blot, Derivative Assay, Two Tailed Test, Transfection, Over Expression, Fluorescence, Stable Transfection

Journal: iScience

Article Title: MALAT1 regulates human macrophage metabolism by interacting with HADHB

doi: 10.1016/j.isci.2026.115107

Figure Lengend Snippet: MALAT1 suppresses and restrains pro-inflammatory phenotype in macrophages (A and B) Time-course gene expression of MALAT1 (A) and HADHB (B) in PMA-differentiated THP-1-derived macrophages following LPS stimulation at the indicated time points. For each LPS stimulation time point, n = 9 independent experiments (independent cell cultures and stimulated on different days); data are shown as mean ± SD and analyzed with one-way ANOVA followed by Tukey’s post hoc test. (C–H) Time-course gene expression of inflammatory cytokines, macrophage-polarization markers, and HADHB in THP-1-derived macrophages transfected with siMALAT1 or siNC and stimulated with LPS at the indicated time points. For each LPS stimulation time point, n = 9 independent experiments (independent cell cultures and stimulated on different days); data are shown as mean ± SD and analyzed with two-way ANOVA followed by Tukey’s post hoc test. (I) Western blot showing HADHB protein levels in THP-1-derived macrophages after siMALAT1 transfection and LPS stimulation at the indicated time points; adjacent quantification shows HADHB band intensity normalized to GAPDH (loading control). The experiment was repeated n = 3 independent experiments (independent cell cultures on different days); data are shown as mean ± SD and analyzed with one-way ANOVA followed by Tukey’s post hoc test.

Article Snippet: Human THP-1 cell lines and Raw 264.7 cell lines were purchased from the American Type Culture Collection (ATCC, TIB-202, TIB-71).

Techniques: Gene Expression, Derivative Assay, Transfection, Western Blot, Control

Journal: iScience

Article Title: MALAT1 regulates human macrophage metabolism by interacting with HADHB

doi: 10.1016/j.isci.2026.115107

Figure Lengend Snippet: MALAT1- HADHB pathway constrains inflammation and attenuates macrophage pro-inflammatory responses (A and B) Temporal expression profiles of IL1B , TNF , TGFB , MRC1 , NOS2 , and HADHB in THP-1-derived macrophages transfected with siHADHB or siNC and stimulated with LPS at the indicated time points. For each LPS stimulation time point, n = 3 independent experiments (independent cell cultures and stimulated on different days); data are shown as mean ± SD and analyzed with one-way ANOVA followed by Tukey’s post hoc test. (C–E) Rescue experiment, Oe- HADHB THP-1-derived macrophages were transfected with siMALAT1; figures show LPS-evoked expression changes in inflammatory, polarization, and HADHB markers relative to negative control (siNC). For each LPS stimulation time point, n = 3 independent experiments (independent cell cultures and stimulated on different days); data are shown as mean ± SD and analyzed with one-way ANOVA followed by Tukey’s post hoc test.

Article Snippet: Human THP-1 cell lines and Raw 264.7 cell lines were purchased from the American Type Culture Collection (ATCC, TIB-202, TIB-71).

Techniques: Expressing, Derivative Assay, Transfection, Negative Control

Journal: iScience

Article Title: MALAT1 regulates human macrophage metabolism by interacting with HADHB

doi: 10.1016/j.isci.2026.115107

Figure Lengend Snippet: MALAT1 orchestrates global transcriptional and metabolic reprogramming in macrophages (A) Differentially expressed genes (DEGs) identified by RNA-seq in LPS-stimulated macrophages after 24 h. The experiment was repeated; n = 3 independent experiments (independent cell cultures on different days). (B and C) Hallmark gene set scores from gene set enrichment analysis (GSEA). The experiment was repeated; n = 3 independent experiments (independent cell cultures on different days). (D and E) Intracellular and extracellular L-lactate levels in PMA-differentiated macrophages transfected with siMALAT1 or siNC and subsequently stimulated with LPS; measurements were taken at the indicated post-LPS time points. For each LPS stimulation time point, n = 3 independent experiments (independent cell cultures and stimulated on different days); data are shown as mean ± SD and analyzed with two-way ANOVA followed by Tukey’s post hoc test. (F, G, J, and L) The gene expression levels of LDHA , LDHB , ACLY , and CD36 in LPS-stimulated siRNA-transfected THP-1-derived macrophages. For each LPS stimulation time point, n = 9 independent experiments (independent cell cultures and stimulated on different days); data are shown as mean ± SD and analyzed with two-way ANOVA followed by Tukey’s post hoc test. (H, I, and K) The protein expression of LDHA, LDHB, ACLY, and p-ACLY in LPS-stimulated THP-1-derived macrophages transfected with siMALAT1 or siNC at the indicated time points. The experiment was repeated; n = 3 independent experiments (independent cell cultures on different days).

Article Snippet: Human THP-1 cell lines and Raw 264.7 cell lines were purchased from the American Type Culture Collection (ATCC, TIB-202, TIB-71).

Techniques: RNA Sequencing, Transfection, Gene Expression, Derivative Assay, Expressing

Journal: iScience

Article Title: MALAT1 regulates human macrophage metabolism by interacting with HADHB

doi: 10.1016/j.isci.2026.115107

Figure Lengend Snippet: Mechanistic illustration of MALAT1 -mediated regulation in THP-1-derived macrophages during inflammation Under inflammatory conditions, MALAT1 expression in the nucleus increases and drives the metabolic reprogramming of macrophages through several mechanisms. First, MALAT1 enhances HADHB expression and trans -localizes from the nucleus to mitochondria via the HuR-MTCH2 axis, where it binds HADHB and elevates its thiolase activity to promote FAO. MALAT1 also upregulates LDHB, facilitating the lactate-to-pyruvate conversion, while downregulating LDHA to reduce lactate production and diminish glycolysis. Furthermore, MALAT1 downregulates ACLY expression and lowers phosphorylated ACLY, thereby inhibiting the fatty acid synthesis pathway typically associated with the pro-inflammatory phenotype.

Article Snippet: Human THP-1 cell lines and Raw 264.7 cell lines were purchased from the American Type Culture Collection (ATCC, TIB-202, TIB-71).

Techniques: Derivative Assay, Expressing, Activity Assay

Journal: bioRxiv

Article Title: Towards molecular-based functional classification of fetal bovine serum

doi: 10.64898/2026.03.16.712020

Figure Lengend Snippet: (A) Hierarchical clustering dendrograms of FBS conditions based on differential gene expression profiles in MRC-5 (left), Jurkat (middle), and THP-1 (right) cells. Clustering was performed using the number of differentially expressed genes (DEGs; |log2FC| > 0.6, adjusted p-value < 0.05) as a distance metric. Horizontal red lines indicate the clustering threshold of 100 DEGs, defining distinct FBS clusters. MRC-5 and THP-1 cells show 4 distinct clusters, while Jurkat cells display 5 clusters, revealing cell type-specific responses to FBS alternatives. (B) Module size distributions from WGCNA for MRC-5 (left), Jurkat (middle), and THP-1 (right) cells. Bar colors represent distinct co-expression modules identified using signed networks with Pearson correlation (soft-thresholding power selected to achieve scale-free topology R ≥ 0.8; minimum module size = 30 genes; merge cut height = 0.25). MRC-5 cells: 7 modules (turquoise: 4,728 genes; blue: 3,580; brown: 2,417; grey: 936; yellow: 634; green: 447; red: 234). Jurkat cells: 7 modules (turquoise: 4,234; blue: 3,635; grey: 2,289; brown: 1,464; yellow: 660; green: 240; red: 204). THP-1 cells: 5 modules (turquoise: 4,201; blue: 4,16; grey: 2,478; brown: 2,184; yellow: 434). (C) Module-trait relationship heatmaps showing Pearson correlations between module eigengenes and FBS cluster assignments for MRC-5 (left), Jurkat (middle), and THP-1 (right) cells. Each cell displays the correlation coefficient (color scale: blue = -0.5, white = 0, red = 0.5) and corresponding p-value. Significant correlations (p < 0.05, calculated using corPvalueStudent) indicate modules with expression patterns specifically associated with FBS clusters. Notable associations include: MRC-5 turquoise module with Cluster 1 (r = - 0.45, p = 3.22×10 ); Jurkat red module with Cluster 2 (r = 0.76, p = 1.38×10 ); THP-1 yellow module with Cluster 1 (r = 0.65, p = 2.54×10 8).

Article Snippet: The adherent human lung fibroblast cell line MRC-5 (CCL-171), the suspension human monocytic cell line THP-1 (TIB-202), and the human acute T-cell leukemic cell line Jurkat Clone E6-1 (TIB-152) were purchased from American Type Culture Collection (ATCC, USA).

Techniques: Gene Expression, Expressing