20536 1 ap  (Proteintech)

Journal: Bioactive Materials

Article Title: Multi-omics-informed hydrogel design: modulating IL-6 to reduce endoplasmic reticulum stress in bone regeneration

doi: 10.1016/j.bioactmat.2025.09.005

Figure Lengend Snippet: Bulk RNA-seq identification of early regulatory genes and signals in bone defects. ( A ) Experimental workflow for bulk RNA-seq in early stages of bone defects in rats. ( B ) Construction of interaction networks and molecular dynamics simulations for key regulatory proteins in early-stage femoral and alveolar bone defects. ( C – E ) Immunohistochemical detection and quantitative analysis of Heat Shock Protein Family A (Hsp70) Member 5 (HSPA5) and interleukin-6 (IL-6) expression levels in early healing tissues of the femoral and alveolar regions. Scale bars: 2 mm, 100 μm, and 50 μm. ( F , G ) Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analysis results for differentially expressed genes (DEGs) in early healing stages of femoral and alveolar bone defects. Statistical significance was determined using a t -test.

Article Snippet: First, HSPA5 (bs-1219R, Bioss, CN) was used, followed by IL-6 (MA5-51298, Thermo Fisher Scientific, US), ARG1 (bsm-56207R, Bioss, CN), FHOD3 (bs-13156R, Bioss, CN), RUNX2 (bs-1134R, Bioss, CN), and Osterix (YA3539, MCE, US).

Techniques: RNA Sequencing, Immunohistochemical staining, Expressing

Journal: Bioactive Materials

Article Title: Multi-omics-informed hydrogel design: modulating IL-6 to reduce endoplasmic reticulum stress in bone regeneration

doi: 10.1016/j.bioactmat.2025.09.005

Figure Lengend Snippet: Protein identification for key proteins and signals in early bone defects. ( A ) Workflow for tissue protein identification at early stages of bone defects in rats. ( B , C ) KEGG and GO enrichment analysis of differential proteins in early healing tissues of femoral and alveolar bone defects. ( D , E ) Interaction network analysis of HSPA5 and IL-6 within the enrichment signals of femoral and alveolar bones. ( F ) Protein expression levels of HSPA5 and IL-6 in early healing tissues of femoral and alveolar defects. ( G ) Quantitative analysis of HSPA5 and IL-6 protein expression. Statistical analysis employed a t -test.

Article Snippet: First, HSPA5 (bs-1219R, Bioss, CN) was used, followed by IL-6 (MA5-51298, Thermo Fisher Scientific, US), ARG1 (bsm-56207R, Bioss, CN), FHOD3 (bs-13156R, Bioss, CN), RUNX2 (bs-1134R, Bioss, CN), and Osterix (YA3539, MCE, US).

Techniques: Expressing

Journal: Bioactive Materials

Article Title: Multi-omics-informed hydrogel design: modulating IL-6 to reduce endoplasmic reticulum stress in bone regeneration

doi: 10.1016/j.bioactmat.2025.09.005

Figure Lengend Snippet: IL-6-mediated regulation of HSPA5 in early bone defect repair tissues. ( A ) Extraction of femoral and alveolar bone cell clusters from whole-cell atlases. ( B ) Pseudo-time series analysis of cellular and gene expression in repair tissues. ( C ) The rat model for femoral and alveolar bone defect treatment using HA15 and LMT28. ( D , E ) Immunohistochemical analysis of HSPA5 and IL-6 expression in femoral defects after HA15 treatment. Scale bars: 200 μm and 50 μm. ( F , G ) Immunohistochemical analysis in alveolar defects after LMT28 treatment. Scale bars: 200 μm and 50 μm. HA15 inhibits HSPA5; LMT28 inhibits IL-6. Statistical significance was assessed with a t -test.

Article Snippet: First, HSPA5 (bs-1219R, Bioss, CN) was used, followed by IL-6 (MA5-51298, Thermo Fisher Scientific, US), ARG1 (bsm-56207R, Bioss, CN), FHOD3 (bs-13156R, Bioss, CN), RUNX2 (bs-1134R, Bioss, CN), and Osterix (YA3539, MCE, US).

Techniques: Extraction, Gene Expression, Immunohistochemical staining, Expressing

Journal: Bioactive Materials

Article Title: Multi-omics-informed hydrogel design: modulating IL-6 to reduce endoplasmic reticulum stress in bone regeneration

doi: 10.1016/j.bioactmat.2025.09.005

Figure Lengend Snippet: M2 macrophages release IL-6 in alveolar bone defect repair. ( A ) Extraction of Il-6 -expressing cell populations. ( B ) Identification and classification of subpopulations within Il-6 -expressing cells. ( C ) Identification of the Il-6 + M2 cell subpopulation. ( D ) Temporal association between Il-6 + M2 cell subpopulation and samples. ( E – G ) Multiplex immunofluorescence for HSPA5 (GRP78/BiP), M2 macrophages (ARG1), and IL-6, combined with statistical analysis of fluorescence intensity and colocalization analysis. Scale bars: 1 mm, 100 μm and 50 μm. ( H ) KEGG and GO enrichment analysis of DEGs in the Il-6 + M2 cell subpopulation.

Article Snippet: First, HSPA5 (bs-1219R, Bioss, CN) was used, followed by IL-6 (MA5-51298, Thermo Fisher Scientific, US), ARG1 (bsm-56207R, Bioss, CN), FHOD3 (bs-13156R, Bioss, CN), RUNX2 (bs-1134R, Bioss, CN), and Osterix (YA3539, MCE, US).

Techniques: Extraction, Expressing, Multiplex Assay, Immunofluorescence, Fluorescence

Journal: Bioactive Materials

Article Title: Multi-omics-informed hydrogel design: modulating IL-6 to reduce endoplasmic reticulum stress in bone regeneration

doi: 10.1016/j.bioactmat.2025.09.005

Figure Lengend Snippet: IL-6 modulates HSPA5 to mitigate endoplasmic reticulum stress (ERS)-related apoptosis in early bone defects. ( A , B ) Expression and quantitative analysis of ERS-related proteins following HA15 and LMT28 treatments. Scale bars: 200 μm and 50 μm. ( C , D ) Quantitative analysis of apoptosis-related proteins CHOP and caspase-12 after treatment. Scale bars: 200 μm and 50 μm. ( E ) Expression levels of ERS and apoptosis-related proteins were analyzed by Western blot, and statistical analysis was conducted on the results. ( F , G ) Quantitative terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining and statistical analysis of apoptosis in femoral and alveolar bone defects. Scale bars: 100 μm. ( H ) Ca 2+ histopathological staining. Scale bars: 100 μm and 40 μm. ( I ) Detection of reactive oxygen species in repair tissue following HA15 and LMT28 treatment. Scale bars: 200 μm and 50 μm. Statistical significance was determined using one-way ANOVA.

Article Snippet: First, HSPA5 (bs-1219R, Bioss, CN) was used, followed by IL-6 (MA5-51298, Thermo Fisher Scientific, US), ARG1 (bsm-56207R, Bioss, CN), FHOD3 (bs-13156R, Bioss, CN), RUNX2 (bs-1134R, Bioss, CN), and Osterix (YA3539, MCE, US).

Techniques: Expressing, Western Blot, TUNEL Assay, Staining

Journal: Bioactive Materials

Article Title: Multi-omics-informed hydrogel design: modulating IL-6 to reduce endoplasmic reticulum stress in bone regeneration

doi: 10.1016/j.bioactmat.2025.09.005

Figure Lengend Snippet: Comparative analysis of the early repair processes of femoral and alveolar defects. ( A ) Overview of the research timeline after preparing rat bone defect models. ( B , C ) Hematoxylin and eosin staining and its quantitative analysis of early repair tissue between femoral and alveolar defects at different time intervals. Scale bars: 2 mm and 100 μm *, bone tissue. ( D ) Immunofluorescence staining of Runt-related transcription factor (Runx2) and Osterix proteins in early-stage repair tissues of femoral and alveolar bone defects at different time intervals. Scale bars: 1 mm and 50 μm. ( E , F ) Western blot detection and statistical analysis of Runx2 and Osterix protein expression in repair tissues from femoral and alveolar bone at 7 days and 13 days. ( G – J ) Micro-CT analysis of femoral and alveolar defects on days 7 and 13. Scale bar: 1 mm. Parameters include bone mineral density (BMD), bone volume fraction (BV/TV), trabecular thickness (Tb.Th), and trabecular separation (Tb.Sp). Statistical significance was assessed using a t -test.

Article Snippet: First, HSPA5 (bs-1219R, Bioss, CN) was used, followed by IL-6 (MA5-51298, Thermo Fisher Scientific, US), ARG1 (bsm-56207R, Bioss, CN), FHOD3 (bs-13156R, Bioss, CN), RUNX2 (bs-1134R, Bioss, CN), and Osterix (YA3539, MCE, US).

Techniques: Staining, Immunofluorescence, Western Blot, Expressing, Micro-CT

Journal: Neural Regeneration Research

Article Title: Chitosan alleviates symptoms of Parkinson’s disease by reducing acetate levels, which decreases inflammation and promotes repair of the intestinal barrier and blood–brain barrier

doi: 10.4103/NRR.NRR-D-23-01511

Figure Lengend Snippet: Chitosan alleviates motor dysfunction and improves DA neuron survival in an MPTP-induced mouse model of PD. (A) Experimental timeline of behavioral tests and sample collection from the different treatment groups, including control, NaA alone, MPTP-induced PD model, MPTP + NaA, chitosan treatment, MPTP + chitosan + PPARD antagonist, and SCFA treatment. (B) Experimental timeline of cell treatment with acetate, an AMPK agonist, and a PPARD agonist. (C) Chitosan significantly increased mouse body weight ( n = 7/group). (D) In the rotarod test, fall latency was increased after chitosan treatment ( n = 7/group). (E) Chitosan administration significantly increased TH expression, as determined by western blot assay. GAPDH was used as loading control ( n = 3/group). (F) Chitosan treatment significantly increased the number of TH-positive dopaminergic neurons (red, Alexa Fluor 594), as determined by immunofluorescence staining ( n = 3/group). Scale bars: 100 μm. (G) UHPLC-MS/MS was used to detect DA, DOPAC, and HVA levels in striatum tissue ( n = 4/group). Treatment with chitosan significantly upregulated the levels of DA, DOPAC/DA, and (DOPAC + HVA)/DA, but there was no significant change in HVA/DA levels. All data are presented as the mean ± SD. All experiments were repeated at least three times. * P < 0.05 (two-way analysis of variance followed by Tukey’s multiple comparisons test (C) or one-way analysis of variance followed by Tukey’s multiple comparisons test (D–G). AMPK: Adenosine 5′-monophosphate-activated protein kinase; DA: dopamine; DOPAC: 3,4-dihydroxyphenylacetic acid; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; HVA: homovanillic acid; ig: intragastrical administration; ip: intraperitoneal administration; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; n.s.: no significance; NaA: sodium acetate; PD: Parkinson’s disease; PPARD: peroxisome proliferator-activated receptor delta; SCFA: short-chain fatty acid; SN: substantia nigra; TH: tyrosine hydroxylase.

Article Snippet: The primary antibodies used were as follows: rabbit anti-glyceraldehyde-3-phosphate dehydrogenase polyclonal antibody (GAPDH; 1:10,000, Proteintech, Wuhan, Hubei, China, Cat# 10494-1-AP, RRID: AB_2263076), rabbit anti-TH polyclonal antibody (1:5000, Proteintech, Cat# 25859-1-AP, RRID: AB_2716568), rabbit anti-zonula occludens-1 polyclonal antibody (ZO-1; 1:5000, Proteintech, Cat# 21773-1-AP, RRID: AB_10733242), rabbit anti-occludin polyclonal antibody (1:15,000, Proteintech, Cat# 27260-1-AP, RRID: AB_2880820), rabbit anti-AMPKα polyclonal antibody (1:1000, Cell Signaling Technology, Danvers, Massachusetts, USA, Cat# 2532, RRID: AB_330331), rabbit anti-phospho-AMPKα monoclonal antibody (1:1000, Cell Signaling Technology, Cat# 2535, RRID: AB_331250), and rabbit anti-PPARD polyclonal antibody (1:1000, Abcam, Cambridge, UK, Cat# ab23673, RRID: AB_2165902).

Techniques: Control, Expressing, Western Blot, Immunofluorescence, Staining, Tandem Mass Spectroscopy

Journal: Neural Regeneration Research

Article Title: Chitosan alleviates symptoms of Parkinson’s disease by reducing acetate levels, which decreases inflammation and promotes repair of the intestinal barrier and blood–brain barrier

doi: 10.4103/NRR.NRR-D-23-01511

Figure Lengend Snippet: The PPAR and AMPK signaling pathways are enriched in genes whose expression is altered by acetate supplementation, as determined by RNA sequencing of colon tissue from a mouse model of PD. (A) Venn diagram of DEGs among three datasets. (B) KEGG pathway enrichment analysis showed that these DEGs were enriched in the PPAR and AMPK signaling pathways, which are associated with inflammation. (C) Heatmap analysis of 183 common DEGs between MPTP vs . MPTP + Chitosan and MPTP + Chitosan vs . MPTP + Chitosan + NaA ( n = 2/group). (D) qPCR verification analysis of the mRNA levels (normalized to the control group) of the PPAR and AMPK signaling pathway–related genes whose expression was altered in mouse colon tissue ( n = 3/group). All data are presented as the mean ± SD. All experiments were repeated three times. * P < 0.05 (one-way analysis of variance followed by Tukey’s multiple comparisons test). AMPK: Adenosine 5′-monophosphate-activated protein kinase; DEGs: differentially expressed genes; FABP5: fatty acid-binding protein 5; FASN: fatty acid synthase; KEGG: Kyoto Encyclopedia of Genes and Genomes; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; n.s.: not significant; NaA: sodium acetate; PD: Parkinson’s disease; PPAR: peroxisome proliferators-activated receptor; PPARD: peroxisome proliferator-activated receptor delta; qPCR: quantitative polymerase chain reaction; SCD1: stearoyl-coenzyme A desaturase 1; SCD4: stearoyl-coenzyme A desaturase 4.

Article Snippet: The primary antibodies used were as follows: rabbit anti-glyceraldehyde-3-phosphate dehydrogenase polyclonal antibody (GAPDH; 1:10,000, Proteintech, Wuhan, Hubei, China, Cat# 10494-1-AP, RRID: AB_2263076), rabbit anti-TH polyclonal antibody (1:5000, Proteintech, Cat# 25859-1-AP, RRID: AB_2716568), rabbit anti-zonula occludens-1 polyclonal antibody (ZO-1; 1:5000, Proteintech, Cat# 21773-1-AP, RRID: AB_10733242), rabbit anti-occludin polyclonal antibody (1:15,000, Proteintech, Cat# 27260-1-AP, RRID: AB_2880820), rabbit anti-AMPKα polyclonal antibody (1:1000, Cell Signaling Technology, Danvers, Massachusetts, USA, Cat# 2532, RRID: AB_330331), rabbit anti-phospho-AMPKα monoclonal antibody (1:1000, Cell Signaling Technology, Cat# 2535, RRID: AB_331250), and rabbit anti-PPARD polyclonal antibody (1:1000, Abcam, Cambridge, UK, Cat# ab23673, RRID: AB_2165902).

Techniques: Protein-Protein interactions, Expressing, RNA Sequencing, Control, Binding Assay, Real-time Polymerase Chain Reaction

Journal: Neural Regeneration Research

Article Title: Chitosan alleviates symptoms of Parkinson’s disease by reducing acetate levels, which decreases inflammation and promotes repair of the intestinal barrier and blood–brain barrier

doi: 10.4103/NRR.NRR-D-23-01511

Figure Lengend Snippet: Acetate may relieve inflammation by activating the PPARD/AMPK signaling pathway in Caco-2 cells. (A, B) Western blot analysis of PPARD, p-AMPK, and AMPK expression in Caco-2 cells treated with a PPARD agonist or left untreated. Compared with the acetate group, PPARD and p-AMPK expression levels were significantly increased in the group treated with acetate and the PPARD agonist. (C) qPCR was used to detect the mRNA level of IL-1β, IL-6, IL-8, IL-10, TNF-α, and iNOS in Caco-2 cells treated with a PPARD agonist or left untreated. Compared with the acetate group, IL-1β and TNF-α were down-regulated, and iNOS was up-regulated, in cells treated with the PPARD agonist. (D, E) Western blot analysis of AMPK, p-AMPK, and PPARD expression levels in Caco-2 cells treated with an AMPK agonist or left untreated. p-AMPK expression was significantly lower in the group treated with acetate and an AMPK agonist than in the acetate-only group. PPARD expression was not altered by treatment with the AMPK agonist. (F) qPCR was used to detect the mRNA levels of IL-1β, IL-6, IL-8, IL-10, TNF-α, and iNOS in Caco-2 cells treated with the AMPK agonist or left untreated. Treatment with the AMPK agonist reduced IL-1β, iNOS, IL-6, and TNF-α expression levels compared with treatment with acetate alone. GAPDH was used as loading control in the western blot assays. All data (normalized by control group) are presented as the mean ± SD ( n = 3/group). All experiments were repeated at least three times. * P < 0.05 (one-way analysis of variance followed by Tukey’s multiple comparisons test). AMPK: Adenosine 5′-monophosphate-activated protein kinase; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; IL-1β: interleukin-1 beta; IL-6: interleukin-6; IL-8: interleukin-8; IL-10: interleukin-10; iNOS: inductible nitric oxide synthase; n.s.: no significance; p-AMPK: phosphorylation adenosine 5’-monophosphate-activated protein kinase; PPARD: peroxisome proliferator-activated receptor delta; qPCR: quantitative polymerase chain reaction; TNF-α: tumor necrosis factor alpha.

Article Snippet: The primary antibodies used were as follows: rabbit anti-glyceraldehyde-3-phosphate dehydrogenase polyclonal antibody (GAPDH; 1:10,000, Proteintech, Wuhan, Hubei, China, Cat# 10494-1-AP, RRID: AB_2263076), rabbit anti-TH polyclonal antibody (1:5000, Proteintech, Cat# 25859-1-AP, RRID: AB_2716568), rabbit anti-zonula occludens-1 polyclonal antibody (ZO-1; 1:5000, Proteintech, Cat# 21773-1-AP, RRID: AB_10733242), rabbit anti-occludin polyclonal antibody (1:15,000, Proteintech, Cat# 27260-1-AP, RRID: AB_2880820), rabbit anti-AMPKα polyclonal antibody (1:1000, Cell Signaling Technology, Danvers, Massachusetts, USA, Cat# 2532, RRID: AB_330331), rabbit anti-phospho-AMPKα monoclonal antibody (1:1000, Cell Signaling Technology, Cat# 2535, RRID: AB_331250), and rabbit anti-PPARD polyclonal antibody (1:1000, Abcam, Cambridge, UK, Cat# ab23673, RRID: AB_2165902).

Techniques: Western Blot, Expressing, Control, Phospho-proteomics, Real-time Polymerase Chain Reaction

Journal: Neural Regeneration Research

Article Title: Chitosan alleviates symptoms of Parkinson’s disease by reducing acetate levels, which decreases inflammation and promotes repair of the intestinal barrier and blood–brain barrier

doi: 10.4103/NRR.NRR-D-23-01511

Figure Lengend Snippet: Chitosan may reduce acetate levels, thereby activating the PPARD-AMPK signaling pathway, which promotes repair of the intestinal barrier and reduces neuroinflammation in an MPTP-induced mouse model of PD. (A, B) Western blot analysis of p-AMPK, AMPK, and PPARD levels in mouse colon tissue ( n = 3/group). Treatment with acetate significantly increased p-AMPK and PPARD expression. (C) Treatment with a PPARD antagonist significantly decreased mouse body weight ( n = 6/group). (D) There were no significant differences in fall latency among the groups in the rotarod test, which was used to assess motor dysfunction ( n = 6/group). (E–G) PPARD antagonist treatment significantly decreased PPARD, TH, ZO-1, and occludin expression, as determined by western blot ( n = 3/group). (H) Immunofluorescence staining for ZO-1 (green, Alexa Fluor 488) and occludin (red, Alexa Fluor 594) in mouse colon tissue ( n = 3/group). The PPARD antagonist treatment group exhibited markedly reduced ZO-1 and occludin mRNA expression levels in colon tissue. Scale bars: 10 μm. (I) QPCR was used to measure the mRNA levels of IL-1β, IL-6, IL-8, IL-10, TNF-α, and iNOS in mouse colon tissue ( n = 3/group). Treatment with the PPARD antagonist increased IL-6 and TNF-α mRNA levels, while IL-8 and iNOS levels were reduced. (J) ELISA was used to detect IL-1β, IL-6, IL-10, and TNF-α expression levels in mouse plasma ( n = 5/group). IL-1β, IL-6, and TNF-α expression levels were significantly increased in the PPARD antagonist treatment group. (K) QPCR was used to measure mRNA levels of IL-1β, IL-6, IL-8, IL-10, TNF-α, and iNOS in the SN ( n = 3/group). Treatment with the PPARD antagonist significantly increased the mRNA levels of IL-1β, IL-6, and IL-8. (L) Treatment with the PPARD antagonist reduced p-AMPK, but not AMPK, expression ( n = 3/group). GAPDH was used as the internal reference. All data are presented as the mean ± SD. All experiments were repeated at least three times. * P < 0.05 (one-way analysis of variance followed by Tukey’s multiple comparisons test (A, B) or unpaired t -test (C–L)). AMPK: Adenosine 5′-monophosphate-activated protein kinase; DAPI: 4′,6-diamidino-2-phenylindole; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; IL-1β: interleukin-1 Beta; IL-6: interleukin-6; IL-8: interleukin-8; IL-10: interleukin-10; iNOS: inductible nitric oxide synthase; n.s.: not significant; NaA: sodium acetate; p-AMPK: phosphorylation adenosine 5′-monophosphate-activated protein kinase; PD: Parkinson’s disease; PPARD: peroxisome proliferator-activated receptor delta; QPCR: quantitative polymerase chain reaction; SN: substantia nigra; TH: tyrosine hydroxylase; TNF-α: tumor necrosis factor alpha; ZO-1: Zonula occludens-1.

Article Snippet: The primary antibodies used were as follows: rabbit anti-glyceraldehyde-3-phosphate dehydrogenase polyclonal antibody (GAPDH; 1:10,000, Proteintech, Wuhan, Hubei, China, Cat# 10494-1-AP, RRID: AB_2263076), rabbit anti-TH polyclonal antibody (1:5000, Proteintech, Cat# 25859-1-AP, RRID: AB_2716568), rabbit anti-zonula occludens-1 polyclonal antibody (ZO-1; 1:5000, Proteintech, Cat# 21773-1-AP, RRID: AB_10733242), rabbit anti-occludin polyclonal antibody (1:15,000, Proteintech, Cat# 27260-1-AP, RRID: AB_2880820), rabbit anti-AMPKα polyclonal antibody (1:1000, Cell Signaling Technology, Danvers, Massachusetts, USA, Cat# 2532, RRID: AB_330331), rabbit anti-phospho-AMPKα monoclonal antibody (1:1000, Cell Signaling Technology, Cat# 2535, RRID: AB_331250), and rabbit anti-PPARD polyclonal antibody (1:1000, Abcam, Cambridge, UK, Cat# ab23673, RRID: AB_2165902).

Techniques: Western Blot, Expressing, Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Phospho-proteomics, Real-time Polymerase Chain Reaction

Journal: Genes & Diseases

Article Title: Bmp9 modulates cell proliferation and intercellular junctions in HERS during tooth root development

doi: 10.1016/j.gendis.2025.101777

Figure Lengend Snippet: Loss of Bmp9 hinders the differentiation and secretion of odontoblasts. (A, B) Hematoxylin-eosin staining of mandibular first molars of wild-type (WT) littermates and Bmp9 knockout (KO) mice on postnatal day 0 (PN0) (A) and at 2 weeks (B) ( n = 7). The dashed-line boxes are enlarged. The asterisks indicate the odontoblasts. (C) Immunohistochemical staining of nestin in the mandibular first molars of WT littermates and Bmp9 -KO mice at 2 weeks of age ( n = 6). The dashed-line boxes are enlarged. The asterisks indicate the odontoblasts. (D) Sirius red staining of mandibular first molars of WT littermates and Bmp9 -KO mice at 2 weeks of age ( n = 6). The dashed-line boxes are enlarged. (E) Immunohistochemical staining of DSPP in the mandibular first molars of WT littermates and Bmp9 -KO mice at 2 weeks of age ( n = 6). The dashed-line boxes are enlarged. The asterisks indicate the odontoblasts. (F, G) Quantitative analysis of nestin and DSPP expression. Scale bars = 100 μm.

Article Snippet: Primary antibodies were applied at 4 °C overnight, including BMP9 (1:100, Thermo Fisher, PA5-11931), DSPP (1:200, Abcam, ab216892), and Nestin (1:100, Bioss, bs-0008R).

Techniques: Staining, Knock-Out, Immunohistochemical staining, Expressing

Journal: Neural Regeneration Research

Article Title: Changes in border-associated macrophages after stroke: Single-cell sequencing analysis

doi: 10.4103/NRR.NRR-D-24-01092

Figure Lengend Snippet: pySCENIC analysis showing distinct transcription factor regulation. (A) Dot plot of the transcription factor activities of Mrc1 macrophage. (B, C) Pseudotime trajectory of macrophages. (D, E) The expression levels of Tnf and Stat3 followed by pseudotime trajectory. Stat3: signal transducer and activator of transcription 3; Tnf: tumor necrosis factor.

Article Snippet: The rabbit anti-STAT3 polyclonal antibody (1:200, Cat# CL488-60199, RRID: AB_2883117), rabbit anti-TNF-α monoclonal antibody (1:200, Cat# CL488-60291, RRID: AB_2883136), rabbit anti-F4/80 polyclonal antibody (1:500, Cat# CL488-28463, UNIPROT ID Q61549 ), rabbit anti-Iba1 monoclonal antibody (1:500, Cat# 81728-1-RR, UNIPROT ID P55008 ), and goat anti-rabbit IgG (H+L)-CoraLite-488 antibody (1:200, Cat# SA00013-2, RRID: AB_2797132) were all purchased from Wuhan Sanying Biotechnology Co., LTD.

Techniques: Expressing

Journal: Neural Regeneration Research

Article Title: Changes in border-associated macrophages after stroke: Single-cell sequencing analysis

doi: 10.4103/NRR.NRR-D-24-01092

Figure Lengend Snippet: BAM depletion protects against acute stroke. (A) Flowchart of the experimental design. (B) Cerebral blood perfusion images of the brain within 60 seconds in the control and treated groups 24 hours after surgery. Less blue color in the right zone of brain in the treated group indicates better blood flow compared with the control group. (C) TTC staining images of brain from the control and treated groups 24 hours after surgery. Less white color was observed in the right brain zone of the treated group compared with the control group. Scale bars: 1 cm. (D) Infarct volume was quantified by TTC staining. (E) Neurological scores were assessed at 24 hours in the control and treated groups. (F) TNF-α, STAT3, and F4/80 immunopositivity in the control and treated groups 24 hours after surgery. (G) ELISA of TNF-α and STAT3 plasma levels 3 days after surgery. Data are expressed as mean ± SD ( n = 4 in both groups). * P < 0.05, ** P < 0.01, *** P < 0.001 (two-tailed Student’s t -test). BAM: Border-associated macrophage; CBF: cerebral blood flow; ELISA: enzyme linked immunosorbent assay; HE: hematoxylin-eosin staining; IF: immunofluorescence staining; STAT3: signal transducer and activator of transcription 3; TNF: tumor necrosis factor; TTC: 2,3, 5-triphenyltetrazolium chloride.

Article Snippet: The rabbit anti-STAT3 polyclonal antibody (1:200, Cat# CL488-60199, RRID: AB_2883117), rabbit anti-TNF-α monoclonal antibody (1:200, Cat# CL488-60291, RRID: AB_2883136), rabbit anti-F4/80 polyclonal antibody (1:500, Cat# CL488-28463, UNIPROT ID Q61549 ), rabbit anti-Iba1 monoclonal antibody (1:500, Cat# 81728-1-RR, UNIPROT ID P55008 ), and goat anti-rabbit IgG (H+L)-CoraLite-488 antibody (1:200, Cat# SA00013-2, RRID: AB_2797132) were all purchased from Wuhan Sanying Biotechnology Co., LTD.

Techniques: Control, Staining, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Two Tailed Test, Immunofluorescence