Journal: Cell communication and signaling : CCS

Article Title: BMX, a specific HDAC8 inhibitor, with TMZ for advanced CRC therapy: a novel synergic effect to elicit p53-, β-catenin- and MGMT-dependent apoptotic cell death.

doi: 10.1186/s12964-022-01007-x

Figure Lengend Snippet: Fig. 1 BMX, TMZ, oxaliplatin (Oxp) and doxorubicin (Dox) combination inhibited cell proliferation in CRC cells. (A) The proliferation of BMX, TMZ, Oxp, Dox, BMX plus TMZ, BMX plus Oxp or BMX plus Dox in HT29, HCT116 and RKO cells with various drug concentration. (B) Colony formation capability assay with different treatments of BMX, TMZ, Oxp, BMX plus TMZ, and BMX plus Oxp in HT29, HCT116 and RKO cells; the colonies were counted for quantification. (C) Cell cycle analysis after 48 h treatment with different concentrations of BMX alone or BMX combined with TMZ in HT29, HCT116, and RKO cells and the proportion of cells in each cell cycle phase. SubG1, cell with polyploid chromosome; > 4 N, polyploid cell. (D) Apoptosis analysis after 48 h treatment with different concentrations of BMX alone or BMX combined with TMZ and the apoptotic rate of cells in HT29, HCT116, and RKO cells. All results are shown as mean ± s.d. from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 versus control (HT29 cells); #p < 0.05, ##p < 0.01, ###p < 0.001 vs. control (HCT116 cells); †p < 0.05, ††p < 0.01, †††p < 0.001 versus control (RKO cells)

Article Snippet: The cell proliferation of CRC cells was measured by CCK8 assay (Targetmol, Shanghai, China).

Techniques: Concentration Assay, Cell Cycle Assay, Control

Journal: Cell communication and signaling : CCS

Article Title: BMX, a specific HDAC8 inhibitor, with TMZ for advanced CRC therapy: a novel synergic effect to elicit p53-, β-catenin- and MGMT-dependent apoptotic cell death.

doi: 10.1186/s12964-022-01007-x

Figure Lengend Snippet: Fig. 5 BMX, VPA, SAHA, TMZ, Oxp, and Dox combination inhibited cell proliferation in CRC cells. (A) HDAC8 mRNA expression levels stimulated with BMX (5 µM), VPA (4 mM), SAHA (2 µM) with or without TMZ were determined using qRT-PCR assays. (B) HT29, HCT116, and RKO cells were treated with BMX with or without TMZ for 48 h. Then, cells were harvested for detection of acetyl-histone H3 (Lys9/Lys14), acetyl-histone H4 (Lys8), and HDAC8 by Western blot analysis. (C) The proliferation of BMX, VPA, SAHA with or without TMZ, Oxp (5 µM) and Dox (1 µM) in HT29, HCT116, and RKO cells with treatment durations were assayed using the CCK-8 method. (D) Colony formation capability assay with different treatments of BMX, VPA, SAHA with or without TMZ, Oxp and Dox in HT29, HCT116, and RKO cells; the colonies were counted for quantification. All results are shown as mean ± s.d. from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 versus control (HT29 cells); #p < 0.05, ##p < 0.01, ###p < 0.001 versus control (HCT116 cells); †p < 0.05, ††p < 0.01, †††p < 0.001 versus control (RKO cells)

Article Snippet: The cell proliferation of CRC cells was measured by CCK8 assay (Targetmol, Shanghai, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Control

Journal: Cell communication and signaling : CCS

Article Title: BMX, a specific HDAC8 inhibitor, with TMZ for advanced CRC therapy: a novel synergic effect to elicit p53-, β-catenin- and MGMT-dependent apoptotic cell death.

doi: 10.1186/s12964-022-01007-x

Figure Lengend Snippet: Fig. 7 Autophagy expression levels stimulated with BMX and PCI-34051 in the presence or absence of TMZ. (A) HT29, HCT116, and RKO cells were treated with BMX and PCI-34051 with or without TMZ for 48 h. Then, cells were harvested for detection of acetyl-histone H3 (Lys9/Lys14), acetyl-histone H4 (Lys8), and HDAC8 by Western blot analysis. (B) The proliferation of BMX and PCI-34051 with or without TMZ in HT29, HCT116, and RKO cells. (C) Colony formation capability assay with different treatments of BMX and PCI-34051 with or without TMZ in HT29, HCT116, and RKO cells; the colonies were counted for quantification. (D, E) Expressions of cleaved caspase 3, cleaved PARP, LC3 and p62 proteins in HT29, HCT116, and RKO cells treated with BMX and PCI-34051 with or without TMZ for 48 h. All results are shown as mean ± s.d. from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 versus control (HT29 cells); #p < 0.05, ##p < 0.01, ###p < 0.001 versus control (HCT116 cells); †p < 0.05, ††p < 0.01, †††p < 0.001 versus control (RKO cells)

Article Snippet: The cell proliferation of CRC cells was measured by CCK8 assay (Targetmol, Shanghai, China).

Techniques: Expressing, Western Blot, Control

Journal: Journal of Extracellular Vesicles

Article Title: Calcified apoptotic vesicles from PROCR + fibroblasts initiate heterotopic ossification

doi: 10.1002/jev2.12425

Figure Lengend Snippet: Single‐cell RNA‐sequencing (scRNA‐seq) analysis identifies novel apoptotic‐preferential PROCR + fibroblasts in the early stage of HO. (a) The workflow depicting the collection and processing of specimens of sham and HO tendons for scRNA‐seq. (b) Dimension reduction presentation (via tSNE) of combined single‐cell transcriptome data from Achilles tendons of rats from the sham and HO groups after 1 and 3 weeks ( n = 14402). Each dot represents a single cell and is labelled with corresponding cell categories and is coloured according to its cell type identity. Clusters were generated using a resolution of 0.2 before subclustering into major cell types according to the Methods. The Seurat 3 R‐Package segregation grouped the cells into 6 distinct cell clusters. (c and d) Quantitative analysis of clusters based on the combined single‐cell transcriptome data in (b). (e) tSNE of fibroblast clusters (F1–F5). (f) Apoptosis score analyses of fibroblasts based on (e). (g) Bioinformatic analysis of PROCR + cell populations based on (e). (h) Representation analysis of GO categories showing different functions for PROCR + cells. (i) Representation analysis of KEGG categories showing different functions for PROCR + cells.

Article Snippet: To inhibit fibroblast apoptosis in the Achilles tendon, Z‐VAD‐FMK (T7020, TargetMol) dissolved in normal saline was injected into rats at 1 μg/rat, three times a week.

Techniques: Single Cell, RNA Sequencing, Generated

Journal: Advanced Science

Article Title: VDLIN: A Deep Learning‐Based Platform for Methylcobalamin‐Inspired Immunomodulatory Compound Screening

doi: 10.1002/advs.202413775

Figure Lengend Snippet: Co7 induces Ifnb1 expression via the TLR4 signaling pathway. A) Volcano plot illustrating the distribution of differentially expressed genes (DEGs) between the Co7 and DMSO treatment groups after 3 h in RAW 264.7 cells. Fold changes are presented as log 2 transformations. Red dots represent DEGs upregulated in the Co7 group. B) Protein‐protein interaction (PPI) network analysis of differentially expressed genes (DEGs) in the Co7‐treated group compared to the DMSO group, revealing significant enrichment in pathways associated with the innate immune response and type I interferon signaling. C) KEGG pathway enrichment analysis of DEGs induced by Co7, highlighting associations with innate immune response pathways. D) Co7 (50 µmol/L) exhibiting strong antiviral effects against VSV in RAW 264.7 and HT29 cells. E) Co7 significantly reduced the inflammatory response induced by LPS, VSV, EMCV, and HSV in RAW 264.7 cells. F) Volcano plot representing the differential gene expression analysis between Co7‐ and LPS‐treated RAW 264.7 cells after 3 h of treatment. Fold changes are displayed as log 2 transformations. Red dots indicate genes upregulated in the Co7 group, while blue dots represent downregulated genes compared to LPS treatment. G) Western blot analysis demonstrating that Co7 inhibited the expression of iNOS and COX2, as well as the phosphorylation of NF‐κB‐P65 at the protein level in RAW 264.7 cells. H) Co7 significantly reduced the mortality rate in mice (n = 10 per group) following LPS challenge (20 mg/kg), compared to the PBS control group. RT‐qPCR data were presented as means ± SEM from three independent experiments. Statistical significance was determined using one‐way ANOVA with Bonferroni's multiple comparisons test (left three panels in E), paired‐samples t‐test (D, right panel of EMCV and HSV in E), or the log‐rank test (H). * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Each inhibitor was dissolved in anhydrous DMSO and diluted to its respective working concentration: C29 (10 μM, S6597, Selleck) served as a TLR2 inhibitor; Procyanidin B1 (30 μM, HY‐N0795, MedChemExpress) acted as a TLR4 inhibitor; MyD88‐IN‐1 (30 μM, HY‐149992, MedChemExpress) was used to inhibit MyD88; Pepinh‐TRIF TFA (30 μM, HY‐P2565, MedChemExpress) functioned as a TRIF inhibitor; and GSK8612 (5 μM, T5540, TargetMol) inhibited TBK1/IKKε.

Techniques: Expressing, Gene Expression, Western Blot, Phospho-proteomics, Control, Quantitative RT-PCR

Journal: Journal of experimental & clinical cancer research : CR

Article Title: Chemotherapy-elicited extracellular vesicle CXCL1 from dying cells promotes triple-negative breast cancer metastasis by activating TAM/PD-L1 signaling.

doi: 10.1186/s13046-024-03050-7

Figure Lengend Snippet: Fig. 4 CXCL1EV-dead induces macrophage M2 polarization by activating PD-L1 expression. A Chemokine array assay was conducted to characterize the differences in chemokine content between EV-dead and EV-alive. An ELISA assay was conducted to compare the relative CXCL1 content in EV-dead and EV-alive. B Changes in M2 phenotype polarization of Raw264.7 macrophages when treated with 10 ng/ml murine CXCL1, 50 μg/ ml EV-dead, 50 μg/ml EV-deadshCXCL1, 5 μg/ml CXCL1 neutralizing antibody (NA), or EV-dead and CXCL1-NA combination for 48 h. C Representative images of Transwell assay. Raw264.7 macrophages were treated as indicated for 48 h and then co-cultured with 4 T1 cells. Scale bar: 200 μm. D–F Expression changes of CXCL1 and PD-L1 in Raw264.7 macrophages when treated as indicated for 48 h. Scale bar: 10 μm. G The results of the flow cytometry assay suggested that 50 μg/ml EV-dead treatment for 48 h induced the M2 polarization of Raw264.7 macrophages by activating PD-L1 expression; n = 3. Data are presented as mean ± SD. *p < 0.05, **p < 0.01

Article Snippet: CXCL1 secretion inhibitor screening To screen the potential CXCL1 secretion inhibitor of 4 T1 cells from the Chemokine Inhibitor Library (L7600, TOPSCIENCE, Shanghai, China), 4 T1 cells were treated with 80 types of chemokine inhibitors (1 μM) for 48 h. Subsequently, the concentration of CXCL1 in cell culture supernatants was detected using the Mouse CXCL1 ELISA Kit.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Transwell Assay, Cell Culture, Flow Cytometry

Journal: Stem Cells International

Article Title: Mesenchymal Stem Cells Inhibit Epithelial-to-Mesenchymal Transition by Modulating the IRE1 α Branch of the Endoplasmic Reticulum Stress Response

doi: 10.1155/2023/4483776

Figure Lengend Snippet: Suppression of ER stress contributed to MSC-mediated amelioration of EMT in A549 cells. (a) A549 cells were treated with 100 μ M TUDCA for different times. The protein expression levels of BiP, ATF6, ATF4, XBP-1s and XBP-1u were measured using western blotting and quantified using densitometry in ImageJ software ( n = 3, one-way ANOVA with Duncan's post hoc test). (b) A549 cells were treated with 10 ng/ml TGF- β 1 in the presence or absence of TUDCA for 72 hr. The protein expression levels of BiP, ATF6, ATF4, XBP-1s and XBP-1u were measured using western blotting and quantified using densitometry in ImageJ software ( n = 4, one-way ANOVA with Duncan's post hoc test). (c) A549 cells were treated with 100 μ M TUDCA for different times. The protein expression levels of E-cadherin and vimentin were measured using western blotting and quantified using densitometry in ImageJ software ( n = 3, one-way ANOVA with Duncan's post hoc test). (d) A549 cells were treated with 10 ng/ml TGF- β 1 in the presence or absence of TUDCA for 72 hr. The protein expression levels of E-cadherin and vimentin were measured using western blotting and quantified using densitometry in ImageJ software ( n = 4, one-way ANOVA with Duncan's post hoc test). The data are shown as the means ± SEMs ( ∗∗ P < 0.01, ∗ P < 0.05 vs. the control group; ## P < 0.05, # P < 0.05 vs. the TGF- β 1 group).

Article Snippet: A549 cells were treated with TGF- β 1 (10 ng/ml, 100-21, PeproTech, Rocky Hill, NJ, USA) for 72 hr with or without pretreatment with the ER stress inhibitor TUDCA (100 μ M, abs816166; Absin, Shanghai, China) or IRE1 α -specific inhibitor 4 μ 8c (10 μ M, T6363, Topscience, Shanghai, China) for 1 hr.

Techniques: Expressing, Western Blot, Software, Control