interferon stimulated genes thp1 bluetm isg  (InvivoGen)

Journal: Cells

Article Title: uPAR-Targeting Cytotoxic Antibody–Drug Conjugates Selectively Deplete Proinflammatory Myeloid Cells for Autoimmune Indications

doi: 10.3390/cells15090803

Figure Lengend Snippet: Internalization of anti-uPAR mAb 2G10 in myeloid cells. ( A ) The integrated fluorescence intensity of pHrodo dye-conjugated isotype control and 2G10 mAbs in monocyte-derived M1 macrophages at different time points. ( B ) The integrated fluorescence intensity of pHrodo dye-conjugated isotype control and 2G10 mAbs at indicated concentrations in LPS/IFNγ-stimulated THP1 cells at different time points. The increased fluorescence intensity of 2G10 mAbs indicates uPAR-mediated internalization. ( C ) A representative image of anti-uPAR mAb internalization at 3 h after mAb treatment; 67 nM of mAbs labeled with AlexaFluor488 (green color) was used for imaging. Hoechst dyes (blue color) were used for staining cell nucleus, and SiR-Lysosome (red color) was used for staining lysosomes. Scale bar: 20 µm. ( D ) Quantification of the intensities of isotype control or 2G10 mAb in lysosomes. ( E ) The Pearson correlation of the co-localization of mAb and lysosome signals.

Article Snippet: Human THP1 cells were purchased from ATCC and cultured in RPMI 1640 medium (Thermo Fisher Scientific; Seattle, WA, USA; A1049101) supplemented with 10% heat-inactivated FBS and 0.05 mM 2-mercaptoethanol (Gibco TM ; Seattle, WA, USA; 21985023).

Techniques: Fluorescence, Control, Derivative Assay, Labeling, Imaging, Staining

Journal: International Journal of Nanomedicine

Article Title: BIRC5 Promoter-Driven Nanodrugs Suppress BIRC5-Positive Cancers Independent of ABCB1 Status and IDO1 Expression

doi: 10.2147/IJN.S588571

Figure Lengend Snippet: Anti-cancer potency of PL-MNP-p BIRC5 /As- BIRC5 and PL-MNP-p BIRC5 /dN-BIRC5 in BIRC5-Expressing cells. ( A ) Western blot analysis of BIRC5 expression in MIA PaCa-2 and NTUB1 cells. ACTA1 served as a loading control to verify equal protein loading. (B) MIA PaCa-2 cells were co-treated with lysosomotropic agent 3 nM BAF and IC 50 concentrations of PL-MNP-p BIRC5 /As- BIRC5 or PL-MNP-p BIRC5 /dN-BIRC5 for 96 h. Cell viability was determined using the MTT assay. (C) MIA PaCa-2 cells were treated with PL-MNP-p BIRC5 /Emp or PL-MNP-p BIRC5 /dN-BIRC5 at their respective IC 50 concentrations for 48 h. BIRC5-CASP3 protein–protein interactions were detected by in situ PLA using anti-BIRC5 and anti-CASP3 antibodies. Scale bars: 10 µm. (D) γ-H2AX expression in MIA PaCa-2 cells treated with IC 50 concentrations of PL-MNP-p BIRC5 /As- BIRC5 or PL-MNP-p BIRC5 /dN-BIRC5 for 48 h, assessed by Western blotting. ACTA1 served as a loading control. Data represent at least three independent experiments. The bar graph represents the quantified band intensities from independent experiments, with γ-H2AX expression levels normalized to the corresponding ACTA1 band intensity for each sample. (E) Western blot analysis of ABCB1 expression in NTUB1 and NTU0.017 cells. ACTA1 served as a loading control to verify equal protein loading. (F) Cell viability of MIA PaCa-2, NTUB1, NTU0.017, SK-OV-3 (with or without 25 ng/mL IFNγ), MCF10A, HMEC-1, WI-38 VA13 subline 2RA, HaCaT, and THP1 cells treated with PL-MNP-p BIRC5 /Emp, PL-MNP-p BIRC5 /As- BIRC5 , or PL-MNP-p BIRC5 /dN-BIRC5 for 96 h, measured by the MTT assay and WST assay (for THP-1 cells). Data represent at least three independent experiments. Bar and curve graphs indicate the mean ± SD. Statistical analyses were performed using one-way ANOVA for panels ( B – D ), and two-way ANOVA for panel ( F ). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 vs. PL-MNP-p BIRC5 /Emp or non-BAF-treated group; n.s., not significant. Statistical symbols are color-coded to match the corresponding experimental groups in the figure.

Article Snippet: THP1 cells were kindly provided by Dr. Tzeng-Horng Leu (Department of Pharmacology, National Cheng Kung University, Tainan, Taiwan) and cultured in RPMI containing 10% FBS and PSG.

Techniques: Expressing, Western Blot, Control, MTT Assay, Protein-Protein interactions, In Situ, WST Assay

Journal: bioRxiv

Article Title: Microbiome-derived hydroxyphenyl propanoates enhance antitumour immunity by potentiating gasdermin D activity in tumour-associated myeloid cells

doi: 10.64898/2026.04.23.720410

Figure Lengend Snippet: a, immune cell types in the TME identified through scRNAseq of CD45 + cells isolated from orthotopically implanted M3-9-M OVA cells; b, number of immune cell types detected in M3-9-M OVA tumours through scRNAseq experiment (n = 2 pooled samples per experimental groups); c, number of significantly impacted genes obtained from pairwise comparisons (Wilcoxon rank sum test); d, circos plot of all significantly impacted genes, obtained from pairwise comparisons of all cell types measured, that overlap with each other through identical or shared pathways; e, bar graph of transcriptional regulators of 3,2-HPP impacted genes; f-m, NF-κB and IRF induction in mouse RAW-Dual™ cells and human THP1-Dual™ cells with different pathogen recognition receptor agonists in the presence of HPP metabolites (HI = heat inactivated, three combined experiments, error bars represent standard error, one-way ANOVA test).

Article Snippet: RAW-DualTM cells (rawd-ismip), THP1-DualTM cells (thpd-nfis), HEK-Blue TM IL-1β cells (hkb-il1bv2; InvivoGen), THP1-Null2 Cells (thp-nullz) and THP1-KO-GSDMD cells (thp-kogsdmdz) were obtained from InvivoGen (San Diego, CA, USA).

Techniques: Isolation

Journal: bioRxiv

Article Title: Microbiome-derived hydroxyphenyl propanoates enhance antitumour immunity by potentiating gasdermin D activity in tumour-associated myeloid cells

doi: 10.64898/2026.04.23.720410

Figure Lengend Snippet: a, NF-κB activity in THP1-Dual™ cells treated with LPS in the presence of vehicle or HPP metabolites for 0-24 hours (two combined experiments, error bars represent standard error); b-e, NF-κB or IRF activity in THP1-Dual™ cells treated with different inducers in the presence or absence of HPP isomers for 16 hours (three combined experiments, error bars represent standard error, one-way ANOVA test).

Article Snippet: RAW-DualTM cells (rawd-ismip), THP1-DualTM cells (thpd-nfis), HEK-Blue TM IL-1β cells (hkb-il1bv2; InvivoGen), THP1-Null2 Cells (thp-nullz) and THP1-KO-GSDMD cells (thp-kogsdmdz) were obtained from InvivoGen (San Diego, CA, USA).

Techniques: Activity Assay

Journal: bioRxiv

Article Title: Microbiome-derived hydroxyphenyl propanoates enhance antitumour immunity by potentiating gasdermin D activity in tumour-associated myeloid cells

doi: 10.64898/2026.04.23.720410

Figure Lengend Snippet: a, representation of the proteins with denaturing curves significantly altered (red) or unaltered (black) by 3,2-HPP treatment of THP1-Dual™ cells, using thermal proteome profiling (two combined experiments, proteomic coverage: 4301, NPARC test); b, protein-protein interaction network of proteins with denaturation curves altered by 3,2-HPP treatment (red) and the transcriptional regulators of 3, 2-HPP impacted genes in M3-9-M tumours (yellow); c, impact of 3,2-HPP treatment of THP1-Dual™ cells on the denaturation curve of GSDMD (two-combined experiments, NPARC test); d, NF-κB induction in THP1-Dual™ reporter cells treated with conditioned media harvested from WT or Gsdmd- KO THP1 cells treated with LPS ± HPP metabolites ± IL-1α and −1β neutralizing antibody (three combined experiments, one-way ANOVA with Bonferroni multiple comparison test); e-h, [secreted IL-1α and −1β] in conditioned media harvested from THP-1 WT or Gsdmd -KO cells, or human PBMCs, cultured for 16 hours in LPS ± HPPs (two combined experiments, one-way ANOVA with Bonferroni multiple comparison test); i-l, [secreted IL-1α and −1β] in conditioned media harvested from THP-1 WT vs. Gsdmd -KO cells, or human PBMCs, cultured for 16 hours in LPS and NG ± HPPs (two combined experiments, one-way ANOVA with Bonferroni multiple comparison test); m, western blot of THP-1 cells treated with LPS ± NG ± HPPs (representative of two experiments).

Article Snippet: RAW-DualTM cells (rawd-ismip), THP1-DualTM cells (thpd-nfis), HEK-Blue TM IL-1β cells (hkb-il1bv2; InvivoGen), THP1-Null2 Cells (thp-nullz) and THP1-KO-GSDMD cells (thp-kogsdmdz) were obtained from InvivoGen (San Diego, CA, USA).

Techniques: Comparison, Cell Culture, Western Blot

Journal: bioRxiv

Article Title: Microbiome-derived hydroxyphenyl propanoates enhance antitumour immunity by potentiating gasdermin D activity in tumour-associated myeloid cells

doi: 10.64898/2026.04.23.720410

Figure Lengend Snippet: a, IL-1β activity in the supernatant of THP1-WT vs. - GSDMD -KO cells treated with LPS±HPPs for 16 hours, measured using HEK-Blue™ IL-1β reporter cells (three combined experiments, error bars represent standard error, one-way ANOVA with Bonferroni test); b, LDH release from THP1-WT vs. – GSDMD -KO cells treated with LPS±HPP for 16 hours (three combined experiments, one-way ANOVA with Bonferroni test); c-d, IL-1β activity in the supernatant of THP1-WT vs. - GSDMD -KO cells © and human PBMCs (d) treated with LPS and NG±HPPs for 16 hours. measured using HEK-Blue™ IL-1β reporter cells (three combined experiments, error bars represent standard error, one-way ANOVA with Bonferroni test); e-f, LDH release from THP1-WT vs. – GSDMD -KO cells € and human PBMCs (f) treated with LPS and NG±HPPs for 16 hours (three combined experiments, one-way ANOVA with Bonferroni test).

Article Snippet: RAW-DualTM cells (rawd-ismip), THP1-DualTM cells (thpd-nfis), HEK-Blue TM IL-1β cells (hkb-il1bv2; InvivoGen), THP1-Null2 Cells (thp-nullz) and THP1-KO-GSDMD cells (thp-kogsdmdz) were obtained from InvivoGen (San Diego, CA, USA).

Techniques: Activity Assay