Journal: Molecular Metabolism

Article Title: Indoleamine 2,3-dioxygenase mediates the therapeutic effects of adipose-derived stromal/stem cells in experimental periodontitis by modulating macrophages through the kynurenine-AhR-NRF2 pathway

doi: 10.1016/j.molmet.2022.101617

Figure Lengend Snippet: ASC therapeutic effects on experimental periodontitis in rats. (A) The schematic diagram showed the induction of the experimental rat model of periodontitis for 21 days and ASC injection after the removal of ligatures. (B) Micro-CT imaging and bone parameters (CEJ-ABC distance, BV/TV, Tb. Th, and Tb. Sp) between healthy and experimental rats of periodontitis. Scale bar, 1 mm. (C) Micro-CT imaging displayed the alveolar bone of PBS-injected and ASC-injected rats on day 7, day 14, and day 21 Scale bar, 1 mm. (D) The CEJ-ABC distance and bone parameters (BV/TV, Tb. Th, and Tb. Sp) between the PBS-injected and ASC-injected rats. (E) Representative immunofluorescence staining of M1 phenotype macrophages (iNOS + ; Green) and M2 macrophages (CD206 + ; Red) in sagittal sections of maxillary molars in PBS-injected and ASC-injected rats. Scale bar, 20 μm. (F) The calculated ratio of iNOS + /CD206 + macrophages in PBS-injected and ASC-injected groups. CEJ-ABC, cementoenamel junction and alveolar bone crest. All data were expressed as mean ± SD; ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: For tissues, the sections were incubated with a mouse anti-CD206 monoclonal antibody (1:100, 60143-1-lg, Proteintech), a rabbit anti-NRF2 monoclonal antibody (1:100, A0674, Abclonal), a rabbit anti-inducible nitric oxide synthase (iNOS) polyclonal antibody (1:100, 18985-1-AP, Proteintech) overnight at 4 °C.

Techniques: Injection, Micro-CT, Imaging, Immunofluorescence, Staining

Journal: Molecular Metabolism

Article Title: Indoleamine 2,3-dioxygenase mediates the therapeutic effects of adipose-derived stromal/stem cells in experimental periodontitis by modulating macrophages through the kynurenine-AhR-NRF2 pathway

doi: 10.1016/j.molmet.2022.101617

Figure Lengend Snippet: ASCs modulated macrophage polarization through NRF2. (A) Representative immunohistochemical staining of NRF2 in the PBS-injected and ASC-injected rats on day 7, day 14, and day 21. Scale bar, 50 μm. (B) Representative immunofluorescence staining of NRF2 (Green) and M2 macrophages (CD206 + ; Red) in sagittal sections of maxillary molars in PBS-injected and ASC-injected rats. Scale bar, 20 μm. (C) Schematic diagram of knocking down NRF2 in macrophages and macrophages cocultured with ASCs and LPS stimulation. (D, E) RT-qPCR and western blotting were used to measure the NRF2 siRNA or siNC in macrophages. (F, G) The mRNA expression and protein levels of M1 markers were increased, while those of M2 markers were decreased after silencing NRF2 in the cocultured groups. R, root; PDL, periodontal ligament; AB, alveolar bone; siRNA, small interfering RNA; NC, negative control. All data were expressed as mean ± SD; ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: For tissues, the sections were incubated with a mouse anti-CD206 monoclonal antibody (1:100, 60143-1-lg, Proteintech), a rabbit anti-NRF2 monoclonal antibody (1:100, A0674, Abclonal), a rabbit anti-inducible nitric oxide synthase (iNOS) polyclonal antibody (1:100, 18985-1-AP, Proteintech) overnight at 4 °C.

Techniques: Immunohistochemical staining, Staining, Injection, Immunofluorescence, Quantitative RT-PCR, Western Blot, Expressing, Small Interfering RNA, Negative Control

Journal: Molecular Metabolism

Article Title: Indoleamine 2,3-dioxygenase mediates the therapeutic effects of adipose-derived stromal/stem cells in experimental periodontitis by modulating macrophages through the kynurenine-AhR-NRF2 pathway

doi: 10.1016/j.molmet.2022.101617

Figure Lengend Snippet: Inhibition of IDO activity reduced the therapeutic potential of ASCs in experimental periodontitis and downregulated NRF2 expression. (A) Three-dimensional reconstruction of maxillary alveolar bone in ASC-injected and 1-MT pretreated ASC-injected rats on day 7, day 14, and day 21. Scale bar, 1 mm. (B) Analysis of bone parameters. (C) Immunohistochemical staining of IL-1β, OCN, and NRF2 in the ASC-injected and 1-MT pretreated ASC-injected rats. Scale bar, 50 μm. (D) Representative immunofluorescence staining of M1 phenotype macrophages (iNOS + ; Green) and M2 macrophages (CD206 + ; Red) in sagittal sections of maxillary molars in ASC-injected and 1-MT pretreated ASC-injected rats. Scale bar, 20 μm. (E) The mean IOD of IL-1β, OCN, and NRF2 and the calculated ratio of iNOS + /CD206 + . IOD, integrated optical density.

Article Snippet: For tissues, the sections were incubated with a mouse anti-CD206 monoclonal antibody (1:100, 60143-1-lg, Proteintech), a rabbit anti-NRF2 monoclonal antibody (1:100, A0674, Abclonal), a rabbit anti-inducible nitric oxide synthase (iNOS) polyclonal antibody (1:100, 18985-1-AP, Proteintech) overnight at 4 °C.

Techniques: Inhibition, Activity Assay, Expressing, Injection, Immunohistochemical staining, Staining, Immunofluorescence

Journal: British Journal of Pharmacology

Article Title: Myeloid‐specific dopamine D 2 receptor signalling controls inflammation in acute pancreatitis via inhibiting M1 macrophage

doi: 10.1111/bph.15026

Figure Lengend Snippet: Myeloid‐specific D2 deletion mediates M1 macrophage polarization in vivo. Two models of acute pancreatitis (AP) were induced in WT C57BL6/J and LysM+/creD2 fl/fl on C57BL6/J background mice in vivo. (a) FCM of TNFα, iNOS, and CD206 expression in CD45+CD11b+F4/80+ macrophages from pancreas of caerulein and LPS‐induced AP model. (b) FCM of TNFα, iNOS, and CD206 expression in CD45+CD11b+F4/80+ macrophages from pancreas of l‐arginine‐induced AP model. n = 6 per group, data were expressed as mean ± SEM. Cae, caerulein; l‐Arg, l‐arginine. ƫ P < .05 LysM+/creD2 fl/fl‐AP versus WT‐AP

Article Snippet: Antibodies against TNFα (60291‐1‐Ig) and CD206 (18704‐1‐AP) were from Proteintech Biotechnology (Wuhan, China).

Techniques: In Vivo, Expressing

Journal: British Journal of Pharmacology

Article Title: Myeloid‐specific dopamine D 2 receptor signalling controls inflammation in acute pancreatitis via inhibiting M1 macrophage

doi: 10.1111/bph.15026

Figure Lengend Snippet: Myeloid‐specific D2 deletion mediates M1 macrophage polarization in vitro. BMDMs were isolated and cultured from WT C57BL6/J and LysM+/creD2 fl/fl mice. M1 macrophages were induced by 100 ng·ml−1 LPS and 10 ng·ml−1 IFN‐γ for 24 hr and were treated with 5‐μM quinpirole at the same time. (a) qRT‐PCR of mRNA levels of inos, Tnfα, Il1β and Il6 in M1‐phenotype BMDMs. BMDMs were isolated from six animals per group, data were expressed as mean ± SEM. (b) FCM of TNFα, iNOS, and CD206 expression in M1‐phenotype BMDMs. BMDMs were isolated from six animals per group, data were expressed as mean ± SEM. *P < .05 WT‐M1 versus WT‐M0; # P < .05 WT‐M1 treated with quinpirole (Quin) versus WT‐M1; + P < .05 LysM+/creD2 fl/fl‐M1 versus LysM+/creD2 fl/fl‐M0; ƫ P < .05 LysM+/creD2 fl/fl ‐M1 versus WT‐M1

Article Snippet: Antibodies against TNFα (60291‐1‐Ig) and CD206 (18704‐1‐AP) were from Proteintech Biotechnology (Wuhan, China).

Techniques: In Vitro, Isolation, Cell Culture, Quantitative RT-PCR, Expressing

Journal: British Journal of Pharmacology

Article Title: Myeloid‐specific dopamine D 2 receptor signalling controls inflammation in acute pancreatitis via inhibiting M1 macrophage

doi: 10.1111/bph.15026

Figure Lengend Snippet: D2 receptor activation inhibits M1 macrophage polarization in vitro. M1 macrophages were induced by 100 ng·ml−1 LPS and 10 ng·ml−1 IFN‐γ and were treated with quinpirole (2.5, 5, and 10 μM) at the same time. (a) mRNA levels of inos, Tnfα, Il1β, and Il6 by qRT‐PCR in LPS and IFN‐γ‐stimulated BMDMs. (b) FCM of TNFα, iNOS and CD206 expression in LPS and IFN‐γ‐stimulated BMDMs. n = 6 per group, data were expressed as mean ± SEM. Scale bar = 50 μm. *P < .05 versus NC; # P < .05 versus LPS and IFN‐γ

Article Snippet: Antibodies against TNFα (60291‐1‐Ig) and CD206 (18704‐1‐AP) were from Proteintech Biotechnology (Wuhan, China).

Techniques: Activation Assay, In Vitro, Quantitative RT-PCR, Expressing

Journal: British Journal of Pharmacology

Article Title: Myeloid‐specific dopamine D 2 receptor signalling controls inflammation in acute pancreatitis via inhibiting M1 macrophage

doi: 10.1111/bph.15026

Figure Lengend Snippet: D2 receptor activation prevents pro‐inflammatory M1 macrophage polarization in the pancreas in vivo. Pancreatic leukocytes from control and acute pancreatitis (AP) mice were isolated and sorted for CD11b+F4/80+ macrophages. mRNA levels of inos, Tnfα, Il1β, and Il6 by qRT‐PCR in CD45+CD11b+F4/80+ cells from pancreatic leukocytes during (a) caerulein and LPS‐induced AP model and (b) l‐arginine‐induced AP model. FCM of TNFα, iNOS, and CD206 expression in CD45+CD11b+F4/80+ cells from pancreatic leukocytes during (c) caerulein and LPS‐induced AP model and (d) l‐arginine‐induced AP model. n = 6 per group, data were expressed as mean ± SEM. *P < .05 versus Ctrl; # P < .05 versus AP

Article Snippet: Antibodies against TNFα (60291‐1‐Ig) and CD206 (18704‐1‐AP) were from Proteintech Biotechnology (Wuhan, China).

Techniques: Activation Assay, In Vivo, Isolation, Quantitative RT-PCR, Expressing

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Integrative proteomics and metabolomics analyses reveals the regulation of autophagy and ferroptosis by RAB10 through Slc37a2/mTOR pathway in breast cancer

doi: 10.1007/s00018-025-05788-5

Figure Lengend Snippet: Bioinformatics analysis of RAB10 highly expressed in multiple cancers. ( A ) RNA sequencing data from 33 cancers in the TCGA database were analyzed using the TIMER2.0 platform. ( B ) RAB10 protein expression levels in tumor and normal tissues were analyzed using the UALCAN database. ( C ) DNA methylation levels of RAB10 across various cancers. ( D ) CNV copy number variation analysis of RAB10 expression in breast cancer. ( E ) Survival curves for overall survival (OS) in breast cancer (222981-s-at). ( F ) RAB10 mRNA expression in breast cancer patients across stages I, II, III, IV, and X. ( G ) ROC curves at 1, 3, and 5 years in the prediction model for breast cancer patients. ( H ) Correlation of RAB10 with multiple glucose metabolism genes. ( I ) LOH heterozygosity analysis of RAB10. ( J ) RNAss tumor stemness score of RAB10 in multiple cancers. ( K ) Correlation of RAB10 expression with infiltration levels of macrophages, neutrophils, CD8 + T cells, CD4 + T cells, and Treg cells. Data are the means ± SD ( n = 3 independent experiments); * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: The primary antibody used was RAB10 (Abclonal, A22746), p62(Proteintech, 18420-1-AP), LC3 (Proteintech, 14600-1-AP), HK2 (Proteintech, 22029-1-AP), GLUT1(Abclonal, A6982), E-cadherin (Proteintech, A11509), snail (Proteintech), CD206 (Proteintech, 18704-1-AP), Arg-1(Cell Signaling Technology, 93668), PCNA (Proteintech, 10205-2-AP).

Techniques: RNA Sequencing, Expressing, DNA Methylation Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Integrative proteomics and metabolomics analyses reveals the regulation of autophagy and ferroptosis by RAB10 through Slc37a2/mTOR pathway in breast cancer

doi: 10.1007/s00018-025-05788-5

Figure Lengend Snippet: RAB10 is highly expressed in cancer tissues of BC patients. ( A ) Representative images of histochemical staining in breast cancer tissues and adjacent normal tissues. ( B , C ) Statistical analysis of RAB10 expression scoring in paired samples from breast cancer patients. ( D ) Statistical analysis of RAB10 expression scores in tumor tissues of breast cancer patients based on TNM stage classification. ( E ) Proportion of TNBC-positive patients with high RAB10 expression. ( F ) Proportion of HER2-positive patients with high RAB10 expression. ( G ) Proportion of ER-positive patients with high RAB10 expression. Data are the means ± SD ( n = 3 independent experiments); * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: The primary antibody used was RAB10 (Abclonal, A22746), p62(Proteintech, 18420-1-AP), LC3 (Proteintech, 14600-1-AP), HK2 (Proteintech, 22029-1-AP), GLUT1(Abclonal, A6982), E-cadherin (Proteintech, A11509), snail (Proteintech), CD206 (Proteintech, 18704-1-AP), Arg-1(Cell Signaling Technology, 93668), PCNA (Proteintech, 10205-2-AP).

Techniques: Staining, Expressing

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Integrative proteomics and metabolomics analyses reveals the regulation of autophagy and ferroptosis by RAB10 through Slc37a2/mTOR pathway in breast cancer

doi: 10.1007/s00018-025-05788-5

Figure Lengend Snippet: In vivo experiments confirm that RAB10 deletion inhibits breast cancer growth. ( A ) Western blot analysis showing knockdown efficiency of RAB10 lentiviral vectors. ( B ) qRT-PCR analysis confirming knockdown efficiency of RAB10 lentiviral vectors. ( C ) Digital images of tumors from each group. ( D ) Tumor volume and weight in each group after dissection. ( E ) Growth curves of body weight and tumor volume over time in different mouse groups. Data are the means ± SD ( n = 3 independent experiments); * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: The primary antibody used was RAB10 (Abclonal, A22746), p62(Proteintech, 18420-1-AP), LC3 (Proteintech, 14600-1-AP), HK2 (Proteintech, 22029-1-AP), GLUT1(Abclonal, A6982), E-cadherin (Proteintech, A11509), snail (Proteintech), CD206 (Proteintech, 18704-1-AP), Arg-1(Cell Signaling Technology, 93668), PCNA (Proteintech, 10205-2-AP).

Techniques: In Vivo, Western Blot, Knockdown, Quantitative RT-PCR, Dissection

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Integrative proteomics and metabolomics analyses reveals the regulation of autophagy and ferroptosis by RAB10 through Slc37a2/mTOR pathway in breast cancer

doi: 10.1007/s00018-025-05788-5

Figure Lengend Snippet: Proteomics and untargeted metabolomics analyses of key molecules and metabolic pathways involved in RAB10 induced malignant progression of breast cancer. ( A ) Number of differential proteins detected in positive and negative ion modes. ( B ) Subcellular localization analysis. ( C ) Hierarchical cluster analysis of differential proteins. ( D ) Enrichment bubble plot of KEGG-significant pathways detected by proteomics. ( E ) GO enrichment analysis for proteomics revealed that molecular function (MF) was focused on protein binding activity. ( F ) Number of metabolites detected in positive and negative ion modes and proportion of identified metabolites in each chemical classification. ( G ) Circle heatmap showing up- and down-regulated differential metabolites. ( H ) Enrichment map of major pathways detected by metabolomics. ( I ) Pathway-pathway network analysis. ( J ) Pathway analysis of upregulated and downregulated metabolites. ( K ) Gene Set Enrichment Analysis (GSEA) of RAB10-related pathways. Data are the means ± SD ( n = 3 independent experiments); * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: The primary antibody used was RAB10 (Abclonal, A22746), p62(Proteintech, 18420-1-AP), LC3 (Proteintech, 14600-1-AP), HK2 (Proteintech, 22029-1-AP), GLUT1(Abclonal, A6982), E-cadherin (Proteintech, A11509), snail (Proteintech), CD206 (Proteintech, 18704-1-AP), Arg-1(Cell Signaling Technology, 93668), PCNA (Proteintech, 10205-2-AP).

Techniques: Protein Binding, Activity Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Integrative proteomics and metabolomics analyses reveals the regulation of autophagy and ferroptosis by RAB10 through Slc37a2/mTOR pathway in breast cancer

doi: 10.1007/s00018-025-05788-5

Figure Lengend Snippet: Knockdown of RAB10 inhibits glucose metabolism and autophagy and induces ferroptosis in 4T-1 cells. ( A ) Western Blot showing down-regulation of HK2 and GLUT1 expression levels after down-regulation of RAB10. ( B ) Kit assay for glucose concentration, lactate production, G6P content, citrate (CA) content and ATP levels down-regulated after sh-RAB10. ( C ) Western Blot showing the expression levels of P62, LC3, HMGCR, and GPX4. ( D , E ) Flow cytometry analysis of ROS and LPO content. ( F ) Changes in mitochondrial membrane potential. (HG) Fluorescence detection of mitochondrial membrane potential changes in both groups. ( H , I ) Morphological changes in autophagic vesicles and mitochondria observed through transmission electron microscopy. ( J ) Changes in GSH content following RAB10 knockdown. Data are the means ± SD ( n = 3 independent experiments); * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: The primary antibody used was RAB10 (Abclonal, A22746), p62(Proteintech, 18420-1-AP), LC3 (Proteintech, 14600-1-AP), HK2 (Proteintech, 22029-1-AP), GLUT1(Abclonal, A6982), E-cadherin (Proteintech, A11509), snail (Proteintech), CD206 (Proteintech, 18704-1-AP), Arg-1(Cell Signaling Technology, 93668), PCNA (Proteintech, 10205-2-AP).

Techniques: Knockdown, Western Blot, Expressing, Concentration Assay, Flow Cytometry, Membrane, Fluorescence, Transmission Assay, Electron Microscopy

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Integrative proteomics and metabolomics analyses reveals the regulation of autophagy and ferroptosis by RAB10 through Slc37a2/mTOR pathway in breast cancer

doi: 10.1007/s00018-025-05788-5

Figure Lengend Snippet: In vivo experimental validation of protein expression changes in tumor tissues after RAB10 knockdown. ( A ) IHC analysis of GLUT1, HK2, p62, LC3, GPX4, HMGCR, CD206, Arg-1, E-cadherin, and Snail expression in control and sh-RAB10 groups (scale bar = 100 μm). ( B ) TUNEL staining of tumor sections from different treatment groups (scale bar = 100 μm). ( C ) Immunofluorescence assay for GPX4, LC3, CD206, and Arg-1 expression levels. Data are the means ± SD ( n = 3 independent experiments); * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: The primary antibody used was RAB10 (Abclonal, A22746), p62(Proteintech, 18420-1-AP), LC3 (Proteintech, 14600-1-AP), HK2 (Proteintech, 22029-1-AP), GLUT1(Abclonal, A6982), E-cadherin (Proteintech, A11509), snail (Proteintech), CD206 (Proteintech, 18704-1-AP), Arg-1(Cell Signaling Technology, 93668), PCNA (Proteintech, 10205-2-AP).

Techniques: In Vivo, Biomarker Discovery, Expressing, Knockdown, Control, TUNEL Assay, Staining, Immunofluorescence

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Integrative proteomics and metabolomics analyses reveals the regulation of autophagy and ferroptosis by RAB10 through Slc37a2/mTOR pathway in breast cancer

doi: 10.1007/s00018-025-05788-5

Figure Lengend Snippet: RAB10 was positively correlated with Slc37a2 and Slc37a2 was associated with poor prognosis of breast cancer. ( A ) Decreased expression of Slc37a2 following RAB10 knockdown. ( B ) Reduced activity of glucose-6-phosphate transmembrane transporter after RAB10 knockdown. ( C ) Correlation analysis of RAB10 and Slc37a2. (D) Kaplan-Meier data analyses show that BC patients with high Slc37a2 expression have worse 5-year overall survival and recurrence-free survival ( P < 0.05). ( E ) Correlation analysis of RAB10, TLR4, and Slc37a2. ( F ) Western blot analysis of Slc37a2 expression following sh-RAB10 and RS 09 TFA treatment. ( G ) TNMplot database shows high Slc37a2 expression in BC patients ( P < 0.01). (H) TCGA data analyses show that high Slc37a2 expression in primary tumor ( P < 0.01).(I) TCGA data analyses show that BC patients with high Slc37a2 expression have worse 5-year overall survival and recurrence-free survival ( P < 0.05). Data are the means ± SD ( n = 3 independent experiments); * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: The primary antibody used was RAB10 (Abclonal, A22746), p62(Proteintech, 18420-1-AP), LC3 (Proteintech, 14600-1-AP), HK2 (Proteintech, 22029-1-AP), GLUT1(Abclonal, A6982), E-cadherin (Proteintech, A11509), snail (Proteintech), CD206 (Proteintech, 18704-1-AP), Arg-1(Cell Signaling Technology, 93668), PCNA (Proteintech, 10205-2-AP).

Techniques: Expressing, Knockdown, Activity Assay, Western Blot

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Integrative proteomics and metabolomics analyses reveals the regulation of autophagy and ferroptosis by RAB10 through Slc37a2/mTOR pathway in breast cancer

doi: 10.1007/s00018-025-05788-5

Figure Lengend Snippet: RAB10 regulates autophagy and ferroptosis in breast cancer cells by regulating metabolism reprogramming

Article Snippet: The primary antibody used was RAB10 (Abclonal, A22746), p62(Proteintech, 18420-1-AP), LC3 (Proteintech, 14600-1-AP), HK2 (Proteintech, 22029-1-AP), GLUT1(Abclonal, A6982), E-cadherin (Proteintech, A11509), snail (Proteintech), CD206 (Proteintech, 18704-1-AP), Arg-1(Cell Signaling Technology, 93668), PCNA (Proteintech, 10205-2-AP).

Techniques: