Journal: Materials today. Bio

Article Title: Precise delivery of doxorubicin and imiquimod through pH-responsive tumor microenvironment-active targeting micelles for chemo- and immunotherapy.

doi: 10.1016/j.mtbio.2022.100482

Figure Lengend Snippet: Fig. 10. Immunostaining of tumor tissues in tumor-bearing mice after treatments. (A) Immunohistochemistry images of tumor sections stained with CD3, CD8, and TNF-α antibodies. The scale bar is 100 μm. (B) Immunofluorescence images of tumor tissues after treatments for 12 days. The scale bar is 50 μm. Blue fluorescence represents the cell nucleus stained with DAPI. Green fluorescence represents the iNOS stained with the iNOS antibody conjugated with FITC.

Article Snippet: After 30 min, the tissue slice was stained with diluted iNOS antibody (Miltenyi Biotec, catalog: 130-116-357) at 4 C overnight.

Techniques: Immunostaining, Immunohistochemistry, Staining

Journal: Scientific Reports

Article Title: High-intensity interval training remodels perineuronal nets in the medial prefrontal cortex to drive microglial polarization and alleviate osteoarthritis pain

doi: 10.1038/s41598-026-40823-w

Figure Lengend Snippet: HIIT reduced PNN accumulation and promoted microglial M2 polarization in the mPFC. (A and B) Triple immunofluorescence staining in the mPFC showing PNN, Iba1, and iNOS ( A ) or PNN, Iba1, and Arg1 ( B ). OA rats showed increased PNN expression and pro-inflammatory (Iba1 + iNOS⁺) microglia, while HIIT reduced PNN levels and promoted anti-inflammatory (Iba1 + Arg1⁺) phenotypes. (C and D) Protein expression of iNOS, Arg1, IL-10, IL-1β, and TNF-α in the mPFC, detected by western blot ( C ) and quantified by bar graphs ( D ) ( n = 6). Statistical methods used: One-way ANOVA. ** p < 0.01 vs. the sham group, ## p < 0.01 vs. the OA group. Data are represented as mean ± SD.

Article Snippet: Equal amounts of protein (30–50 μg) were separated on 10–12% SDS-PAGE gels (FuturePAGE, ACE Biotechnology, ET15420LGel), transferred to PVDF membranes (Millipore, Billerica, MA, USA, IPVH00010), blocked with 5% skim milk (BD Biosciences, Franklin Lakes, NJ, USA) for 1 h, and incubated overnight at 4 °C with primary antibodies against β-actin (1:5000, Servicebio, GB15003-100), COL2A1 (1:1000, Santa Cruz Biotechnology, sc-52658), MMP13 (1:1000, Proteintech, 18165-1-AP), iNOS (1:1000, Santa Cruz Biotechnology, sc-7271), Arg1 (1:1000, Santa Cruz Biotechnology, sc-47715), IL-10 (1:1000, Santa Cruz Biotechnology, sc-32815), IL-1β (1:1000, Santa Cruz Biotechnology, sc-12742), and TNF-α (1:1000, Santa Cruz Biotechnology, sc-52746).

Techniques: Immunofluorescence, Staining, Expressing, Western Blot

Journal: Scientific Reports

Article Title: High-intensity interval training remodels perineuronal nets in the medial prefrontal cortex to drive microglial polarization and alleviate osteoarthritis pain

doi: 10.1038/s41598-026-40823-w

Figure Lengend Snippet: PNNs degradation and HIIT alleviate neuroinflammation in the mPFC of OA rats. ( A - D ) Representative triple immunofluorescence staining images of the mPFC, showing the distribution patterns of PNNs, Iba1 (microglial marker), and iNOS ( A ) or Arg1 ( C ). Quantification of panel A is shown in ( B ), and quantification of panel C is shown in ( D ) ( n = 6). ( E and F ) Western blot analysis and corresponding quantification of inflammatory markers, including iNOS, Arg1, IL-1β, TNF-α, and IL-10 in mPFC tissue lysates ( n = 6). Statistical methods used: One-way ANOVA. ** p < 0.01 vs. the OA+Vehicle group, ns not significant vs. the OA+ChABC group.Data are presented as mean ± SD.

Article Snippet: Equal amounts of protein (30–50 μg) were separated on 10–12% SDS-PAGE gels (FuturePAGE, ACE Biotechnology, ET15420LGel), transferred to PVDF membranes (Millipore, Billerica, MA, USA, IPVH00010), blocked with 5% skim milk (BD Biosciences, Franklin Lakes, NJ, USA) for 1 h, and incubated overnight at 4 °C with primary antibodies against β-actin (1:5000, Servicebio, GB15003-100), COL2A1 (1:1000, Santa Cruz Biotechnology, sc-52658), MMP13 (1:1000, Proteintech, 18165-1-AP), iNOS (1:1000, Santa Cruz Biotechnology, sc-7271), Arg1 (1:1000, Santa Cruz Biotechnology, sc-47715), IL-10 (1:1000, Santa Cruz Biotechnology, sc-32815), IL-1β (1:1000, Santa Cruz Biotechnology, sc-12742), and TNF-α (1:1000, Santa Cruz Biotechnology, sc-52746).

Techniques: Immunofluorescence, Staining, Marker, Western Blot

Journal: Scientific Reports

Article Title: High-intensity interval training remodels perineuronal nets in the medial prefrontal cortex to drive microglial polarization and alleviate osteoarthritis pain

doi: 10.1038/s41598-026-40823-w

Figure Lengend Snippet: PNN remodeling precedes microglial polarization. ( A - D ) Representative triple immunofluorescence staining images of the mPFC, showing PNN, Iba1, and iNOS ( A ), or PNN, Iba1, and Arg1 ( C ). Quantification of panel A is shown in ( B ), and quantification of panel C is shown in ( D ) ( n = 6). Statistical methods used: One-way ANOVA. ** p < 0.01. Data are presented as mean ± SD.

Article Snippet: Equal amounts of protein (30–50 μg) were separated on 10–12% SDS-PAGE gels (FuturePAGE, ACE Biotechnology, ET15420LGel), transferred to PVDF membranes (Millipore, Billerica, MA, USA, IPVH00010), blocked with 5% skim milk (BD Biosciences, Franklin Lakes, NJ, USA) for 1 h, and incubated overnight at 4 °C with primary antibodies against β-actin (1:5000, Servicebio, GB15003-100), COL2A1 (1:1000, Santa Cruz Biotechnology, sc-52658), MMP13 (1:1000, Proteintech, 18165-1-AP), iNOS (1:1000, Santa Cruz Biotechnology, sc-7271), Arg1 (1:1000, Santa Cruz Biotechnology, sc-47715), IL-10 (1:1000, Santa Cruz Biotechnology, sc-32815), IL-1β (1:1000, Santa Cruz Biotechnology, sc-12742), and TNF-α (1:1000, Santa Cruz Biotechnology, sc-52746).

Techniques: Immunofluorescence, Staining

Journal: Oncotarget

Article Title: Subanesthetic isoflurane relieves zymosan-induced neutrophil inflammatory response by targeting NMDA glutamate receptor and Toll-like receptor 2 signaling

doi: 10.18632/oncotarget.9091

Figure Lengend Snippet: A. Neutrophils were treated with increasing concentrations of zymosan for indicated time periods, and were subjected to Western blot analysis. B. Neutrophils were incubated with zymosan for 15 min, post-treated with 0.7% isoflurane for 15 min, followed by a continuous incubation with zymosan for a total of 8 h before Western blot analysis. β-actin was used as an inner control for whole cell lysates. C. Neutrophils were co-transfected with or without an iNOS-luc reporter plasmid and indicated siRNAs. The promoter activity of iNOS was determined in the cell lysates after zymosan treatment for 6 h. D. , E. Neutrophils were pretreated with NAI (D) or transfected with NF-κB p65 siRNA (E), followed by incubation with zymosan for 8 h. The nuclear or whole cell lysates were prepared for Western blot analysis. Lamin B was used as an inner control for the lysates of the nuclear fraction. F. Neutrophils were treated as described in (D), followed by incubation with zymosan for 6 h prior to RT-PCR analysis. G. Neutrophils were treated as described in (D), followed by measurement of NO and ONOO − production. H. Neutrophils were treated as described in (B), and the nuclear lysates were prepared for Western blot analysis. Data are represented as the mean ± SEM of 3 replicates or representative of 3 independent experiments. * P < 0.05, # P < 0.01, as compared with the cells exposed to vehicle alone (A). * P < 0.05, # P < 0.01, as compared with the cells exposed to zymosan alone (B-H), or zymosan + scrambled siRNA (E).

Article Snippet: The small interfering RNA (siRNA) duplexes corresponding to mouse TLR2 (sc-40257), TLR4 (sc-40261), MyD88 (sc-35987), c-Src (sc-29859), p47 phox (sc-151963), p38 MAPK (sc-29434), NF-κB p65 (sc-29411), iNOS (sc-36092), NMDA receptor subunit NR1 (sc-36082), and scrambled siRNA (sc-44230) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, Texas, USA).

Techniques: Western Blot, Incubation, Control, Transfection, Plasmid Preparation, Activity Assay, Reverse Transcription Polymerase Chain Reaction

Journal: Oncotarget

Article Title: Subanesthetic isoflurane relieves zymosan-induced neutrophil inflammatory response by targeting NMDA glutamate receptor and Toll-like receptor 2 signaling

doi: 10.18632/oncotarget.9091

Figure Lengend Snippet: Zymosan induces ROS production through a TLR2/MyD88/c-Src/NADPH oxidase pathway, which in turn caused the activation of p38 MAPK and consequently the activation and nuclear translocation of NF-κB. NF-κB thus switches on iNOS expression, and increases NO production and ONOO − release from neutrophils, which eventually permeabilizes adjacent PMVECs. Subanesthetic isoflurane exerts a protective role for this pathological process by inhibiting c-Src activity, which is probably attributed to isoflurane targeting of the NMDA glutamate receptor and thereby suppression of the calcium signaling in neutrophils.

Article Snippet: The small interfering RNA (siRNA) duplexes corresponding to mouse TLR2 (sc-40257), TLR4 (sc-40261), MyD88 (sc-35987), c-Src (sc-29859), p47 phox (sc-151963), p38 MAPK (sc-29434), NF-κB p65 (sc-29411), iNOS (sc-36092), NMDA receptor subunit NR1 (sc-36082), and scrambled siRNA (sc-44230) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, Texas, USA).

Techniques: Activation Assay, Translocation Assay, Expressing, Activity Assay