Journal: Acta Pharmaceutica Sinica. B

Article Title: Boosting 5-ALA-based photodynamic therapy by a liposomal nanomedicine through intracellular iron ion regulation

doi: 10.1016/j.apsb.2021.03.017

Figure Lengend Snippet: Intracellular iron ion regulating mediated PpIX accumulation and DNA repair enzyme inhibition. (A) The membrance fusion of CLs@DiO and MFLs@DiO (scale bar = 10 μm), the distribution of MFLs@DiI in B16–F10 cells (B) (scale bar = 10 μm). (C) The detection of Fe 2+ after incubating with B16–F10 cells for 4 h (scale bar = 25 μm). (D, E) The transformation of 5-ALA analyzed after incubating with B16–F10 cells for 4 h by CLSM (scale bar = 10 μm) and flow cytometry, respectively. (F) DFO inhibits ALKBH2 repair of m1A in dsDNA by using the DpnII digestion assay and semi-quantitative analysis (G), 1: maker, 2: dsDNA (m1A), 3: dsDNA (m1A)+DpnII, 4: dsDNA (m1A)+ALKBH2+Fe 2+ +DFO + DpnII, 5: dsDNA (m1A)+ALKBH2+Fe 2+ +DpnII. Data are presented as means ± SD ( n = 3). ∗∗∗∗ P < 0.0001, ∗∗ P < 0.01, ∗ P < 0.05.

Article Snippet: The B16–F10 cells were seeded into confocal dish (2 × 10 5 cells per dish) for 24 h of incubation and then treated with control, 5-ALA, DFO, 5-ALA + DFO, CLs@5-ALA/DFO, MFLs@5-ALA/DFO (with the same dose of 5-ALA and DFO: 1 mmol/L and 100 μmol/L respectively) for 4 h. After that, FerroOrange working solution (1 mL, 1 μmol/L) was added to each dish and incubated for a certain period of time, and then the samples were observed directly by a confocal laser-scanning microscopy (CLSM, Leica TCS SP8).

Techniques: Enzyme Inhibition Assay, Transformation Assay, Flow Cytometry

Journal: Acta Pharmaceutica Sinica. B

Article Title: Boosting 5-ALA-based photodynamic therapy by a liposomal nanomedicine through intracellular iron ion regulation

doi: 10.1016/j.apsb.2021.03.017

Figure Lengend Snippet: In vitro enhanced PDT of MFLs@5-ALA/DFO. (A) The production of reactive oxygen species (ROS) after 532 nm laser irradiation by fluorescence microscope (scale bar = 200 μm). (B, C) The analyzed of DNA damage by western blotting and comet assay (scale bar = 200 μm). (D) Cells cytotoxicity of 5-ALA, 5-ALA + DFO, MFLs@5-ALA/DFO. (E) Living and dead cell staining and apoptosis test by Calcein-AM and Propidium Iodide (Calcein-AM/PI) (scale bar = 200 μm). (F) Annexin V/PI staining assays of B16–F10 cells. All of them were incubated with different preparations for 4 h and irradiated with 532 nm laser. Data are presented as means ± SD ( n = 3). ∗∗∗ P < 0.001, ∗∗ P < 0.01, ∗ P < 0.05.

Article Snippet: The B16–F10 cells were seeded into confocal dish (2 × 10 5 cells per dish) for 24 h of incubation and then treated with control, 5-ALA, DFO, 5-ALA + DFO, CLs@5-ALA/DFO, MFLs@5-ALA/DFO (with the same dose of 5-ALA and DFO: 1 mmol/L and 100 μmol/L respectively) for 4 h. After that, FerroOrange working solution (1 mL, 1 μmol/L) was added to each dish and incubated for a certain period of time, and then the samples were observed directly by a confocal laser-scanning microscopy (CLSM, Leica TCS SP8).

Techniques: In Vitro, Irradiation, Fluorescence, Microscopy, Western Blot, Single Cell Gel Electrophoresis, Staining, Incubation

Journal: bioRxiv

Article Title: Ligand stabilization enables physiological GITR signaling and antitumor immunity

doi: 10.64898/2026.05.04.722444

Figure Lengend Snippet: KROS-101 suppresses tumor growth and improves survival in hGITR/hGITRL double KI syngeneic mouse tumor models. a , Experimental design for in vivo efficacy studies. Mice bearing established tumors were treated with KROS-101 beginning on day 3 after tumor implantation. Tumor growth, survival, IVIS bioluminescence imaging for B16-F10-Luc2, and immune profiling by flow cytometry and imaging mass cytometry (IMC) were performed at the indicated time points. b,c , Kaplan-Meier survival curves ( b ) and tumor growth kinetics ( c ) in the MC38 colon carcinoma model following treatment with PBS or KROS-101. KROS-101 significantly improved survival and reduced tumor growth. d,e , Survival ( d ) and tumor volume ( e ) of mice bearing YUMM1.7 melanoma tumors treated with PBS or KROS-101. f , Kaplan-Meier survival analysis in the B16-F10-Luc2 melanoma model demonstrating prolonged survival following KROS-101 treatment. g , Representative IVIS bioluminescence images of B16-F10-Luc2 tumors at baseline (day 3) and after treatment (day 10 and day 17), showing reduced tumor burden in KROS-101 treated mice. h , Tumor growth curves for B16-F10-Luc2 tumors following treatment with PBS or KROS-101. Survival curves were compared using the log-rank (Mantel-Cox) test. Tumor volumes are shown as mean ± s.e.m. * p < 0.05; ** p < 0.01.

Article Snippet: MC38 (murine colon adenocarcinoma), B16-F10-Luc2 (luciferase-expressing murine melanoma), and YUMM1.7 (murine melanoma) cells were obtained from ATCC and maintained in DMEM supplemented with 10% FBS and 1% penicillin-streptomycin at 37°C in 5% CO2.

Techniques: In Vivo, Tumor Implantation, Imaging, Flow Cytometry, Mass Cytometry

Journal: bioRxiv

Article Title: Ligand stabilization enables physiological GITR signaling and antitumor immunity

doi: 10.64898/2026.05.04.722444

Figure Lengend Snippet: Experimental design and immune profiling of tumor-bearing mice treated with KROS-101. a , Schematic of the in vivo experimental workflow. Mice were injected with B16-F10-Luc2 or MC38 tumor cells (100,000 cells in 100 μl), followed by IVIS imaging. Mice were randomized and treated with KROS-101 (20 mg kg⁻¹) or PBS on days 7, 9, 11 and 13. Tumors, blood and spleens were collected for downstream analyses. b , Representative IVIS bioluminescence images of B16-F10-Luc2 tumor-bearing mice (used for flow analysis) before and after treatment with PBS or KROS-101, demonstrating reduced tumor signal in KROS-101-treated mice. c , Tumor growth curves showing reduced tumor volume in KROS-101–treated mice compared with PBS controls over the course of treatment. d , Flow cytometry gating strategy used for immune profiling of tumor-infiltrating leukocytes. Sequential gating identifies major immune populations, including lymphocytes, CD45⁺ leukocytes, NK cells, macrophages, neutrophils, CD3⁺ T cells, CD4⁺ and CD8⁺ T cells, regulatory T cells, and functional markers including perforin, granzyme B, PD-1, TNF-α, IFN-γ, TIGIT and TIM-3.

Article Snippet: MC38 (murine colon adenocarcinoma), B16-F10-Luc2 (luciferase-expressing murine melanoma), and YUMM1.7 (murine melanoma) cells were obtained from ATCC and maintained in DMEM supplemented with 10% FBS and 1% penicillin-streptomycin at 37°C in 5% CO2.

Techniques: In Vivo, Injection, Imaging, Flow Cytometry, Functional Assay

Journal: bioRxiv

Article Title: Ligand stabilization enables physiological GITR signaling and antitumor immunity

doi: 10.64898/2026.05.04.722444

Figure Lengend Snippet: KROS-101 remodels the tumor immune microenvironment by enhancing effector CD8⁺ T cell responses and reducing Treg-mediated suppression. a-d , Immune composition and phenotype of TILs in the MC38 model following treatment with PBS or KROS-101. KROS-101 increased the abundance of CD8⁺ effector T cells and CD8:Treg ratios ( a ), increased frequency of effector memory (CD44 + CD62L − ) CD8+ T cells ( b ), and enhanced CD8⁺ effector function, including GITR expression and production of TNF-α, IFN-γ, and granzyme B ( c ), and reduced the frequency of exhausted PD-1⁺TIM-3⁺ CD8⁺ T cells ( d ). e-g , Representative flow cytometry gating strategies for CD4⁺ and CD8⁺ T cells ( e ), Tregs ( f ), and GITR⁺ CD8 T cells ( g ). h , Expression of suppressive markers in tumor-infiltrating Tregs, showing reduced LAP and IL-10 expression following KROS-101 treatment. i-k , Effects of KROS-101 on tumor-infiltrating APCs. KROS-101 increased M1 macrophage and cDC1 frequencies ( i ) and enhanced GITRL expression on cDC1 ( j, k ), accompanied by increased expression of activation and antigen-presentation markers, including CD86 and MHCII on cDC1 cells ( k ). l-r , Immune profiling of B16-F10-Luc2 tumors showing increased immune infiltration following KROS-101 treatment, including higher frequencies of CD45⁺ leukocytes ( l ), CD3⁺ T cells ( m ), and CD8⁺ T cells ( n ). KROS-101 enhanced IFN-γ production by CD8⁺ T cells ( o ), with no significant change in TNF-α ( p ), and reduced expression of inhibitory receptors TIGIT ( q ) and TIM-3 ( r ). s-u , Effects of KROS-101 on Tregs in B16-F10-Luc2 tumors, showing reduced Treg frequency ( s ), decreased LAP expression ( t ), and minimal change in IL-10 expression ( u ). v-y , Effects on DC populations and activation status. KROS-101 increased cDC1 abundance ( v ) without affecting cDC2 frequency ( w ) and enhanced antigen presentation capacity as indicated by increased MHCII ( x ) and CD86 ( y ) expression on cDC1 cells. Data are presented as mean ± s.e.m. Statistical comparisons were performed using two-tailed unpaired Student’s t-test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns, not significant. Each dot represents one biological replicate (individual mouse).

Article Snippet: MC38 (murine colon adenocarcinoma), B16-F10-Luc2 (luciferase-expressing murine melanoma), and YUMM1.7 (murine melanoma) cells were obtained from ATCC and maintained in DMEM supplemented with 10% FBS and 1% penicillin-streptomycin at 37°C in 5% CO2.

Techniques: Expressing, Flow Cytometry, Activation Assay, Immunopeptidomics, Two Tailed Test

Journal: Applications of Nanomaterials in Human Health

Article Title: Nanomaterials and Their Negative Effects on Human Health

doi: 10.1007/978-981-15-4802-4_13

Figure Lengend Snippet: Impact of nanomaterials on organs/cells/tissues

Article Snippet: 1. , Diaminobutane core (DAB) based generation 3, 4, and 5 dendrimer (diaminobutyric polypropylenimine) for gene delivery , Diaminobutyric polypropylenimine (3-, 4-, and 5-), methoxy PEG (~2 kDa) succinimidyl carboxymethyl esters , Bioware PC-3 M-luc-C6 human prostate adenocarcinoma, bioware B16F10-Luc cells, A431 human epidermoid carcinoma, T98G human glioblastoma, DU145 human prostate carcinoma , – , PEGylated G3 and G4-DAB reduced cytotoxicity of dendrimer. G4-DAB with PEG (2 and 5 kDa) reduced toxicity of dendrimer at lower dose of 20 μg/ml , Somani et al. ( ) .

Techniques: In Vitro, In Vivo, Synthesized, Animal Model, Concentration Assay, Activity Assay, Expressing, Blocking Assay

Journal: International Journal of Molecular Sciences

Article Title: Endothelial Mitochondria Transfer to Melanoma Induces M2-Type Macrophage Polarization and Promotes Tumor Growth by the Nrf2/HO-1-Mediated Pathway

doi: 10.3390/ijms25031857

Figure Lengend Snippet: Transplanted HUVEC mitochondria upregulated the expression of antioxidant and mitochondria biogenesis proteins in B16F10 cells. ( A ) Western blot analysis demonstrating the protein expression levels of Nrf2, HO-1, PGC-1α, Sirt1, and LC3B in B16F10 cells with or without HUVEC mitochondrial transplantation at 24 and 48 h. ( B ) Quantitative analysis of Nrf2, HO-1, PGC-1α, Sirt1, and LC3B expression levels in B16F10 cells after the transplantation of HUVEC mitochondria, as determined through Western blotting at 24 and 48 h. Data are presented as the mean ± standard error after ≥3 independent experiments. Statistical significance was assessed using Student’s t test: * p < 0.05, ** p < 0.01, *** p < 0.005.

Article Snippet: B16F10 melanoma cells, purchased from Merck (Darmstadt, Germany), were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (Gibco, Waltham, MA, USA) supplemented with 10% FBS.

Techniques: Expressing, Western Blot, Transplantation Assay

Journal: International Journal of Molecular Sciences

Article Title: Endothelial Mitochondria Transfer to Melanoma Induces M2-Type Macrophage Polarization and Promotes Tumor Growth by the Nrf2/HO-1-Mediated Pathway

doi: 10.3390/ijms25031857

Figure Lengend Snippet: Transplanted HUVEC mitochondria-activated proliferative signaling and inhibited apoptosis in B16F10 cells. ( A ) Western blot analysis demonstrating the activation of the ERK and AKT signaling pathways at 1, 2, and 8 h following HUVEC mitochondrial transplantation. ( B ) Quantitative analysis of the phosphorylation ratios of ERK and AKT after treatment with HUVEC mitochondria, as determined through Western blotting. ( C ) Expression levels of cyclin D1 and cyclin E in B16F10 cells treated with HUVEC mitochondria for 48 h, as determined through Western blotting. ( D ) Quantitative analysis of cyclin D1 expression and cyclin E expression. ( E ) Viability of B16F10 cells at 24 and 48 h after treatment, as evaluated using the CCK8 assay. ( F ) Caspase-3 and cleaved caspase-3 levels in B16F10 cells, analyzed through Western blotting at 24 and 48 h after treatment with HUVEC mitochondria. ( G ) Quantification of the cleaved caspase-3-caspase-3 expression ratio. Data are presented as the mean ± standard error of the mean after ≥3 independent experiments. Statistical significance was assessed using a two-tailed Student’s t -test: * p < 0.05, ** p < 0.01, *** p < 0.005.

Article Snippet: B16F10 melanoma cells, purchased from Merck (Darmstadt, Germany), were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (Gibco, Waltham, MA, USA) supplemented with 10% FBS.

Techniques: Western Blot, Activation Assay, Protein-Protein interactions, Transplantation Assay, Phospho-proteomics, Expressing, CCK-8 Assay, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Endothelial Mitochondria Transfer to Melanoma Induces M2-Type Macrophage Polarization and Promotes Tumor Growth by the Nrf2/HO-1-Mediated Pathway

doi: 10.3390/ijms25031857

Figure Lengend Snippet: Effects of AI-1 and brusatol on B16F10 Cells. ( A ) Western blot analysis comparing the ratio of p-AKT/AKT in B16F10 cells treated with AI-1 (10 μ) and brusatol (40 nM) at 48 h. ( B ) Quantitative analysis of the p-AKT/AKT ratio after treatment with AI-1 and brusatol, as determined through Western blotting at 48 h. ( C ) Western blot analysis comparing the ratio of p-ERK/ERK in B16F10 cells treated with AI-1 (10 μ) and brusatol (40 nM) at 48 h. ( D ) Quantitative analysis of the p-ERK/ERK ratio after treatment with AI-1 and brusatol, as measured through Western blotting at 48 h. ( E ) Cellular ROS levels of B16F10 cells treated with AI-1 (10 μ) and brusatol (40 nM) at 48 h, as determined through flow cytometry. ( F ) Quantification of ROS levels in B16F10 cells treated with AI-1 (10 µM) and brusatol (40 nM) at 48 h, as determined through flow cytometry. ( G ) Evaluation of caspase activity in B16F10 cells treated with AI-1 (10 µM) and brusatol (40 nM) at 48 h, as determined through flow cytometry. ( H ) Quantitative analysis of caspase activity in B16F10 cells treated with AI-1 (10 µ) and brusatol (40 nM) at 48 h, as determined through flow cytometry. Data are presented as the mean ± standard error of the mean after ≥3 independent experiments. Statistical significance was assessed using a two-tailed Student’s t -test: ** p < 0.01, *** p < 0.005.

Article Snippet: B16F10 melanoma cells, purchased from Merck (Darmstadt, Germany), were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (Gibco, Waltham, MA, USA) supplemented with 10% FBS.

Techniques: Western Blot, Flow Cytometry, Activity Assay, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Endothelial Mitochondria Transfer to Melanoma Induces M2-Type Macrophage Polarization and Promotes Tumor Growth by the Nrf2/HO-1-Mediated Pathway

doi: 10.3390/ijms25031857

Figure Lengend Snippet: Xenograft analysis of melanoma tumor growth. ( A ) Mice were inoculated with B16F10 cells that underwent HUVEC mitochondrial transplantation and were monitored for 10 days. ( B ) Tumor size analysis in six mice. ( C ) Western blot analysis comparing the protein expression levels of Nrf2, HO-1, PGC-1α, Sirt1, and LC3B in B16F10 tumors with or without HUVEC mitochondrial transplantation at 10 days. ( D ) Quantification of the protein expression levels of Nrf2, HO-1, PGC-1α, Sirt1, and LC3B in B16F10 tumors with or without HUVEC mitochondrial transplantation at 10 days. ( E ) Western blot analysis of AKT or ERK signaling in B16F10 tumors with or without HUVEC mitochondrial transplantation at 10 days. ( F ) Quantitative analysis of p-AKT/AKT and p-ERK/ERK ratios in B16F10 tumors with or without HUVEC mitochondrial transplantation at 10 days. Data are presented as the mean ± standard error of the mean after ≥3 independent experiments. Statistical significance was assessed using a two-tailed Student’s t -test: * p < 0.05, *** p < 0.005.

Article Snippet: B16F10 melanoma cells, purchased from Merck (Darmstadt, Germany), were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (Gibco, Waltham, MA, USA) supplemented with 10% FBS.

Techniques: Transplantation Assay, Western Blot, Expressing, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Endothelial Mitochondria Transfer to Melanoma Induces M2-Type Macrophage Polarization and Promotes Tumor Growth by the Nrf2/HO-1-Mediated Pathway

doi: 10.3390/ijms25031857

Figure Lengend Snippet: Xenograft analysis of melanoma tumor growth. ( A ) Western blot analysis comparing the protein expression levels of MMP-9 and TGF-β1 in B16F10 tumors with or without HUVEC mitochondrial transplantation at 10 days. ( B ) Quantitative analysis of MMP-9 and TGF-β1 protein expression levels in B16F10 tumors with or without HUVEC mitochondrial transplantation at 10 days. ( C ) Western blot analysis comparing the protein expression levels of MMP-9 and TGF-β1 in B16F10 cells with and without HUVEC mitochondrial transplantation at 24 or 48 h. ( D ) Quantification of MMP-9 and TGF-β1 expression after treatment with HUVEC mitochondria, as measured through Western blotting at 24 or 48 h. ( E ) Western blot analysis comparing iNOS and Arg1 protein levels in B16F10 tumors with or without HUVEC mitochondrial transplantation at 10 days. ( F ) Quantitative analysis of iNOS and MMP-9 protein levels in B16F10 tumors with or without HUVEC mitochondrial transplantation at 10 days. ( G ) Western blot analysis comparing iNOS and Arg1 protein expression levels in RAW264.7 cells treated with or without TGF-β1 (20 ng/mL) at 48 h. ( H ) Quantification of iNOS and MMP-9 protein levels in RAW264.7 cells treated with or without TGF-β1 (20 ng/mL) at 48 h. Data are presented as the mean ± standard error of the mean after ≥3 independent experiments. Statistical significance was assessed using a two-tailed Student’s t -test: * p < 0.05, ** p < 0.01, *** p < 0.005.

Article Snippet: B16F10 melanoma cells, purchased from Merck (Darmstadt, Germany), were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (Gibco, Waltham, MA, USA) supplemented with 10% FBS.

Techniques: Western Blot, Expressing, Transplantation Assay, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Endothelial Mitochondria Transfer to Melanoma Induces M2-Type Macrophage Polarization and Promotes Tumor Growth by the Nrf2/HO-1-Mediated Pathway

doi: 10.3390/ijms25031857

Figure Lengend Snippet: Effects of SB431540 and brusatol on B16F10 Cells. ( A ) Western blot analysis comparing Nrf2 and TGF-β1 protein levels in B16F10 cells with or without SB431540 (10 μM) treatment at 48 h. ( B ) Quantification of Nrf2 and TGF-β1 expression levels after treatment with or without SB431540, measured through Western blotting at 48 h. ( C ) Western blot analysis comparing Nrf2 and TGF-β1 in B16F10 cells treated with or without brusatol (40 nM) at 48 h. ( D ) Quantification of Nrf2 and TGF-β1 expression levels at 48 h after treatment with or without brusatol, as determined through Western blotting. Data are presented as the mean ± standard error of the mean after ≥3 independent experiments. Statistical significance was assessed using a two-tailed Student’s t -test: ** p < 0.01.

Article Snippet: B16F10 melanoma cells, purchased from Merck (Darmstadt, Germany), were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (Gibco, Waltham, MA, USA) supplemented with 10% FBS.

Techniques: Western Blot, Expressing, Two Tailed Test