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