human gingival fibroblast 1  (ATCC)

Journal: Frontiers in Immunology

Article Title: Danthron as a novel IL-6R agonist promotes thrombopoiesis via the SRC/RAS/MAPK pathway

doi: 10.3389/fimmu.2026.1730028

Figure Lengend Snippet: Danthron demonstrates no significant toxicity to Meg-01 and K562 cells in vitro . (A-B) LDH assays were conducted on days 1, 3, and 5 during danthron (2, 4, 8 and 10 μM) intervention in Meg01 and K562 cells. Ctrl (max) is the LDH maximum release control group. (C-D) The proliferation results of Meg-01 and K562 cells treated with danthron (2, 4 and 8 μM) for 1, 3 and 5 Days. The absorbance values were normalized to the corresponding control group at each time point (see Materials and Methods 2.3 for the specific normalization procedure). From day 1 to day 5, the number of viable cells in the Meg-01 and K562 control groups increased by 3.1-fold and 2.89-fold, respectively. (E-F) Cell apoptosis was analyzed by flow cytometry in Meg-01 and K562 cells treated with danthron (2, 4 and 8 μM). (G) The representative images of each group with different concentrations of danthron (2, 4 and 8 μM) on the 5th day. A preliminary quantitative assessment of megakaryocyte-like large cells based on morphological observations. PMA (1.25 nM) is positive control. Scar bar: 100 µm. All data are expressed as mean ± SD. n = 3 per group. * P < 0.05, ** P < 0.01, and *** P < 0.001, vs. the Ctrl group.

Article Snippet: Meg-01 and K562 cells were obtained from the American Type Culture Collection (Bethesda, MD, United States), which were cultured in RPMI-1640 complete medium (Gibco, Thermo Fisher Scientific, Waltham, MA, United States) supplemented with 10% fetal bovine serum (FBS, CAT: SP002030100, Sperikon Biotechnology Co., LTD, Sichuan, China) and 1% (v:v) Penicillin-Streptomycin Solution (Sperikon Biotechnology Co., LTD,Sichuan, China).

Techniques: In Vitro, Control, Flow Cytometry, Positive Control

Journal: Frontiers in Immunology

Article Title: Danthron as a novel IL-6R agonist promotes thrombopoiesis via the SRC/RAS/MAPK pathway

doi: 10.3389/fimmu.2026.1730028

Figure Lengend Snippet: Danthron promotes MK differentiation and maturation in vitro. (A) Giemsa staining images of Meg-01 and K562 cells treated with danthron (2, 4 and 8 μM) and PMA (1.25 nM) on day 5. Scar bar: 25 µm. (B) Phalloidin staining reveals the expression of F-actin and multinuclear formation in each group after danthron (2, 4 and 8 μM) intervention on the 5th day of the two cells. Scar bar: 100 µm. (C) Analysis of DNA ploidy by flow cytometry. The histogram illustrates DNA ploidy. (D) The results of flow cytometry show the expression of CD41 and CD42b on the 5th day of the two cells with danthron (2, 4 and 8 μM) and PMA (1.25 nM) compared with the control group. All data are expressed as mean ± SD. n = 3 per group. * P < 0.05, ** P < 0.01, and *** P < 0.001, vs. the Ctrl group.

Article Snippet: Meg-01 and K562 cells were obtained from the American Type Culture Collection (Bethesda, MD, United States), which were cultured in RPMI-1640 complete medium (Gibco, Thermo Fisher Scientific, Waltham, MA, United States) supplemented with 10% fetal bovine serum (FBS, CAT: SP002030100, Sperikon Biotechnology Co., LTD, Sichuan, China) and 1% (v:v) Penicillin-Streptomycin Solution (Sperikon Biotechnology Co., LTD,Sichuan, China).

Techniques: In Vitro, Staining, Expressing, Flow Cytometry, Control

Journal: Frontiers in Immunology

Article Title: Danthron as a novel IL-6R agonist promotes thrombopoiesis via the SRC/RAS/MAPK pathway

doi: 10.3389/fimmu.2026.1730028

Figure Lengend Snippet: Danthron enhances the expression of transcription factors critical for regulating MK differentiation. (A) Western blot analysis of the expression of transcription factors related to the regulation of MK differentiation after 5 days of danthron treatment of Meg-01 cells. (B) Immunofluorescence images of transcription factors NF-E2 and RUNX1 on day 5 of danthron intervention in Meg-01 cells. Scar bar: 100 µm. All data are expressed as mean ± SD. n = 3 per group. * P < 0.05, ** P < 0.01, vs Ctrl.

Article Snippet: Meg-01 and K562 cells were obtained from the American Type Culture Collection (Bethesda, MD, United States), which were cultured in RPMI-1640 complete medium (Gibco, Thermo Fisher Scientific, Waltham, MA, United States) supplemented with 10% fetal bovine serum (FBS, CAT: SP002030100, Sperikon Biotechnology Co., LTD, Sichuan, China) and 1% (v:v) Penicillin-Streptomycin Solution (Sperikon Biotechnology Co., LTD,Sichuan, China).

Techniques: Expressing, Western Blot, Immunofluorescence

Journal: Frontiers in Immunology

Article Title: Danthron as a novel IL-6R agonist promotes thrombopoiesis via the SRC/RAS/MAPK pathway

doi: 10.3389/fimmu.2026.1730028

Figure Lengend Snippet: Targets and signaling pathway predicted for danthron treatment of thrombocytopenia using network pharmacology and molecular docking. (A) Common targets of danthron and thrombocytopenia Venn diagram (left), danthron-target-Thrombocytopenia network constructed by Cytoscape_v3.7.1 software and the protein-protein interaction (PPI) network of danthron with core targets of thrombocytopenia based on screening conditions of Degree ≥ 22, BC ≥ 0.0053, CC ≥ 0.5 (middle and right). (B) Visualization of cellular components, physiological processes and biofunctional enrichment analysis of potential therapeutic targets of danthron. (C) The top 10 MF terms with the most enriched danthron-related processes are arranged in ascending order of P value. (D) Top 20 molecular mechanisms of KEGG enrichment in thrombocytopenia treated with danthron. The pathway’s gene enrichment is represented by the size of the bubbles, while the range of P value is indicated by their color. (E) The molecular docking demonstrates the capability of danthron to bind to the core target IL-6R. (F) The detection of IL-6R, p-SRC, RAS, p-MEK and p-ERK1/2 in Meg-01 treated with no danthron and treated with danthron for 5 days by western blot. All data are expressed as mean ± SD. n = 3 per group. * P < 0.05, ** P < 0.01, and *** P < 0.001, vs. the Ctrl group.

Article Snippet: Meg-01 and K562 cells were obtained from the American Type Culture Collection (Bethesda, MD, United States), which were cultured in RPMI-1640 complete medium (Gibco, Thermo Fisher Scientific, Waltham, MA, United States) supplemented with 10% fetal bovine serum (FBS, CAT: SP002030100, Sperikon Biotechnology Co., LTD, Sichuan, China) and 1% (v:v) Penicillin-Streptomycin Solution (Sperikon Biotechnology Co., LTD,Sichuan, China).

Techniques: Construct, Software, Biomarker Discovery, Western Blot

Journal: Frontiers in Immunology

Article Title: Danthron as a novel IL-6R agonist promotes thrombopoiesis via the SRC/RAS/MAPK pathway

doi: 10.3389/fimmu.2026.1730028

Figure Lengend Snippet: Danthron promotes MK differentiation and thrombopoiesis via IL-6R/SRC/RAS/MAPK signaling. (A, B) The flow cytometry analysis the expression of CD41 and CD42b on the 5th day of Meg-01 cells. The histogram illustrates the percentage of CD41 + /CD42b + cells. All data are expressed as mean ± SD. n = 3 per group. * P < 0.05, ** P < 0.01, *** P < 0.001, vs danthron. (C, E) The representative images of Meg-01 of each group with different treatments on the 5th day. Scar bar: 100 µm, n = 3 per group. (D, F) Giemsa staining images of Meg-01 with different treatments on the 5th day. Scar bar: 25 µm, n = 3 per group. (G) Related pathway proteins expression of each group. All data are expressed as mean ± SD. n = 3 per group. * P < 0.05, ** P < 0.01, *** P < 0.001, vs danthron.

Article Snippet: Meg-01 and K562 cells were obtained from the American Type Culture Collection (Bethesda, MD, United States), which were cultured in RPMI-1640 complete medium (Gibco, Thermo Fisher Scientific, Waltham, MA, United States) supplemented with 10% fetal bovine serum (FBS, CAT: SP002030100, Sperikon Biotechnology Co., LTD, Sichuan, China) and 1% (v:v) Penicillin-Streptomycin Solution (Sperikon Biotechnology Co., LTD,Sichuan, China).

Techniques: Flow Cytometry, Expressing, Staining

Journal: PLOS Pathogens

Article Title: Platelet-like particles released from Ebola virus-infected megakaryocytic cells behave like virus-like particles

doi: 10.1371/journal.ppat.1013985

Figure Lengend Snippet: A. Percentage of GFP-positive MEG-01 VP30 cells. PMA-treated or untreated cells were exposed to EBOVΔVP30-GFP at an MOI of 5 and GFP-positive cells were quantified by flow cytometry 2 days after exposure. Data are representative of two independent experiments. B. EBOVΔVP30 titers from MEG-01 VP30 cells. PMA-treated or untreated cells were exposed to EBOVΔVP30-GFP at an MOI of 1. Supernatants were collected daily, and viral titers were determined using Vero VP30 cells. Data are presented as means ± SD of three independent experiments. The dotted lines indicate the lower limit of detection (20 ffu/ml). Statistical significance was assessed by using the multiple unpaired t-test. * p < 0.05, ** p < 0.01.

Article Snippet: Wild-type MEG-01 cells (ATCC, CRL-2021), MEG-01 VP30 cells (MEG-01 cell line stably expressing EBOV VP30), MEG-01 CD41-mCherry cells (MEG-01 cell line stably expressing CD41 [GenBank accession no. NM_000419.5 ] fused with mCherry at the C-terminus [CD41-mCherry]), and MEG-01 CD41-mCherry/GP cells (MEG-01 cell line stably expressing CD41-mCherry and EBOV GP) were cultured in RPMI-1640 medium containing 10% fetal bovine serum (FBS) and antibiotics.

Techniques: Flow Cytometry

Journal: PLOS Pathogens

Article Title: Platelet-like particles released from Ebola virus-infected megakaryocytic cells behave like virus-like particles

doi: 10.1371/journal.ppat.1013985

Figure Lengend Snippet: A. Detection of EBOV proteins by western blot from PLPs released form EBOVΔVP30-exposed MEG-01 VP30 cells. PLPs were collected on day 4 post-exposure from PMA-treated MEG-01 WT or VP30 cells exposed to EBOVΔVP30 at an MOI of 5. The indicated proteins were analyzed by immunoblotting. Data are representative of two independent experiments. B. Relative amount of EBOV gRNA in PLPs released from EBOVΔVP30-exposed MEG-01 VP30 cells. PLPs were prepared as described in (A). EBOV gRNA was quantified by RT-qPCR using the indicated genome-specific primer pairs and normalized to the gRNA in PLPs from MEG-01 WT cells. Data are presented as means ± SD from two independent experiments performed in triplicate. C. Localization of EBOV GP in PLPs released from EBOVΔVP30-exposed MEG-01 VP30 cells. PLPs were prepared as described in (A). The platelet marker CD41 (magenta) and EBOV GP (green) were visualized by immunofluorescence microscopy using specific antibodies, along with the corresponding bright-field image. Scale bars, 5 μm. D. Interaction between EBOV NP/VP35 (top panel) and NP/VP40 (bottom panel) in PLPs released from EBOVΔVP30-exposed MEG-01 VP30 cells. PLPs were prepared as described in (A). NP/VP35 and NP/VP40 complexes (green) were visualized by using a proximity ligation assay (PLA) with specific antibodies and overlaid on the corresponding bright-field image. Scale bars, 5 μm.

Article Snippet: Wild-type MEG-01 cells (ATCC, CRL-2021), MEG-01 VP30 cells (MEG-01 cell line stably expressing EBOV VP30), MEG-01 CD41-mCherry cells (MEG-01 cell line stably expressing CD41 [GenBank accession no. NM_000419.5 ] fused with mCherry at the C-terminus [CD41-mCherry]), and MEG-01 CD41-mCherry/GP cells (MEG-01 cell line stably expressing CD41-mCherry and EBOV GP) were cultured in RPMI-1640 medium containing 10% fetal bovine serum (FBS) and antibiotics.

Techniques: Western Blot, Quantitative RT-PCR, Marker, Immunofluorescence, Microscopy, Proximity Ligation Assay

Journal: PLOS Pathogens

Article Title: Platelet-like particles released from Ebola virus-infected megakaryocytic cells behave like virus-like particles

doi: 10.1371/journal.ppat.1013985

Figure Lengend Snippet: Relative expression levels of EBOV mRNA (A) and gRNA (B) in PLPs released from EBOVΔVP30-exposed MEG-01 VP30 cells. PLPs were collected on day 4 post-exposure from PMA-treated MEG-01 VP30 cells exposed to EBOVΔVP30-GFP at an MOI of 5. EBOV mRNA and gRNA were quantified by RT-qPCR using the indicated specific primer pairs and normalized to the amount in the day 0 samples. Data are presented as means ± SD of three independent experiments performed in triplicate.

Article Snippet: Wild-type MEG-01 cells (ATCC, CRL-2021), MEG-01 VP30 cells (MEG-01 cell line stably expressing EBOV VP30), MEG-01 CD41-mCherry cells (MEG-01 cell line stably expressing CD41 [GenBank accession no. NM_000419.5 ] fused with mCherry at the C-terminus [CD41-mCherry]), and MEG-01 CD41-mCherry/GP cells (MEG-01 cell line stably expressing CD41-mCherry and EBOV GP) were cultured in RPMI-1640 medium containing 10% fetal bovine serum (FBS) and antibiotics.

Techniques: Expressing, Quantitative RT-PCR

Journal: PLOS Pathogens

Article Title: Platelet-like particles released from Ebola virus-infected megakaryocytic cells behave like virus-like particles

doi: 10.1371/journal.ppat.1013985

Figure Lengend Snippet: A,B. Expression levels of GP and CD41-mCherry in MEG-01 cells stably expressing CD41-mCherry with or without EBOV GP (A), and in PLPs released from these stable cell lines (B). Protein levels were analyzed by immunoblotting using the indicated antibodies. C. Percent of mCherry-positive cells that internalized PLPs containing CD41-mCherry or CD41-mCherry/GP. Huh7 VP30 cells were co-incubated with the indicated PLPs for 1, 3, or 5 h, followed by quantification of mCherry-positive cells by flow cytometry. Data are presented as means ± SD of three independent experiments. Statistical significance was assessed by use of a two-way ANOVA followed by Turkey’s multiple comparisons test. * p < 0.05, **** p < 0.0001.

Article Snippet: Wild-type MEG-01 cells (ATCC, CRL-2021), MEG-01 VP30 cells (MEG-01 cell line stably expressing EBOV VP30), MEG-01 CD41-mCherry cells (MEG-01 cell line stably expressing CD41 [GenBank accession no. NM_000419.5 ] fused with mCherry at the C-terminus [CD41-mCherry]), and MEG-01 CD41-mCherry/GP cells (MEG-01 cell line stably expressing CD41-mCherry and EBOV GP) were cultured in RPMI-1640 medium containing 10% fetal bovine serum (FBS) and antibiotics.

Techniques: Expressing, Stable Transfection, Western Blot, Incubation, Flow Cytometry

Journal: PLOS Pathogens

Article Title: Platelet-like particles released from Ebola virus-infected megakaryocytic cells behave like virus-like particles

doi: 10.1371/journal.ppat.1013985

Figure Lengend Snippet: Number of GFP-positive cells following co-incubation with PLPs released from EBOVΔVP30-exposed MEG-01 cells (A). PLPs were collected from PMA-treated MEG-01 WT and VP30 cells exposed to EBOVΔVP30-GFP. Huh7 VP30 cells (2 x 10 5 cells) were co-incubated with the indicated PLPs (2 x 10 6 ) for two days. As a control, cells were cultured in the final wash supernatant (B). GFP-positive cells were quantified by flow cytometry. Data are representative of two independent experiments. C-E. EBOVΔVP30 titers in three different VP30-expressing cell types co-cultured with PLPs released from EBOVΔVP30-exposed MEG-01 VP30 cells. PLPs were prepared as described in (A). Huh7 VP30 cells (2 x 10 5 cells, C), HUVEC VP30 cells (2 x 10 5 cells, D), and PMA-differentiated THP-1 VP30 cells (2 x 10 5 cells, E) were co-cultured with the indicated PLPs (2 x 10 6 ). Virus titers on days 2, 4, and 6 were determined using Vero VP30 cells. Data are presented as means ± SD of three independent experiments. The dotted lines indicate the lower limit of detection (1.3 log10 ffu/ml). Statistical significance was assessed by use of a one-way ANOVA followed by Turkey’s multiple comparisons test. *** p < 0.001, **** p < 0.0001.

Article Snippet: Wild-type MEG-01 cells (ATCC, CRL-2021), MEG-01 VP30 cells (MEG-01 cell line stably expressing EBOV VP30), MEG-01 CD41-mCherry cells (MEG-01 cell line stably expressing CD41 [GenBank accession no. NM_000419.5 ] fused with mCherry at the C-terminus [CD41-mCherry]), and MEG-01 CD41-mCherry/GP cells (MEG-01 cell line stably expressing CD41-mCherry and EBOV GP) were cultured in RPMI-1640 medium containing 10% fetal bovine serum (FBS) and antibiotics.

Techniques: Incubation, Control, Cell Culture, Flow Cytometry, Expressing, Virus