anti nos2  (Bioss)

Journal: Bioactive Materials

Article Title: Immunomodulatory effects of biodegradable Mg–Cu–Zn alloy in esophageal cancer

doi: 10.1016/j.bioactmat.2026.02.046

Figure Lengend Snippet: Distribution of CD163 + M2 TAMs in AKR-derived allograft tumor tissues from immunocompetent C57BL/6 mice. (a, b) Representative IHC staining images showing CD163 + M2 TAMs in the (a) peritumoral stroma and (b) tumor islets. Lower panels display higher-magnification views of the regions outlined by red dashed boxes. (c, d) Quantification of CD163 + cells in the (c) peritumoral stroma and (d) tumor islets. (e) Comparison of CD163 + cell density between the peritumoral stroma and tumor islets. (f) Total number of CD163 + cells in allograft tumors (peritumoral stroma and tumor islets combined). (g, h) Comparison of the density between iNOS + cells and CD163 + cells in the (g) peritumoral stroma and (h) tumor islets. (i, j) Quantification of iNOS + /CD163 + ratio in the (i) peritumoral stroma and (j) tumor islets. p < 0.05 (∗), p < 0.01 (∗∗), p < 0.001 (∗∗∗). A field of view is ∼0.086 mm 2 in (c−j).

Article Snippet: Tissue sections were then incubated with primary antibodies against iNOS (22226-1-AP, ProteinTech, China), CD163 (A26411PM, Abclone, China), CD8 (SP16, Maixin, China), CD4 (SP35, Maixin, China) or Ki-67 (12202S, Cell Signaling Technology) for 12 h at 4 °C, followed by secondary antibodies (Beyotime Biotechnology, Nantong, China).

Techniques: Derivative Assay, Immunohistochemistry, Comparison

Journal: Bioactive Materials

Article Title: Immunomodulatory effects of biodegradable Mg–Cu–Zn alloy in esophageal cancer

doi: 10.1016/j.bioactmat.2026.02.046

Figure Lengend Snippet: Distribution of CD163 + M2 TAMs in AKR-derived allograft tumor tissues from immunocompetent C57BL/6 mice. (a, b) Representative IHC staining images showing CD163 + M2 TAMs in the (a) peritumoral stroma and (b) tumor islets. Lower panels display higher-magnification views of the regions outlined by red dashed boxes. (c, d) Quantification of CD163 + cells in the (c) peritumoral stroma and (d) tumor islets. (e) Comparison of CD163 + cell density between the peritumoral stroma and tumor islets. (f) Total number of CD163 + cells in allograft tumors (peritumoral stroma and tumor islets combined). (g, h) Comparison of the density between iNOS + cells and CD163 + cells in the (g) peritumoral stroma and (h) tumor islets. (i, j) Quantification of iNOS + /CD163 + ratio in the (i) peritumoral stroma and (j) tumor islets. p < 0.05 (∗), p < 0.01 (∗∗), p < 0.001 (∗∗∗). A field of view is ∼0.086 mm 2 in (c−j).

Article Snippet: Tissue sections were then incubated with primary antibodies against iNOS (22226-1-AP, ProteinTech, China), CD163 (A26411PM, Abclone, China), CD8 (SP16, Maixin, China), CD4 (SP35, Maixin, China) or Ki-67 (12202S, Cell Signaling Technology) for 12 h at 4 °C, followed by secondary antibodies (Beyotime Biotechnology, Nantong, China).

Techniques: Derivative Assay, Immunohistochemistry, Comparison

Journal: Journal of Advanced Research

Article Title: Tussilagone attenuated cigarette smoke-induced chronic obstructive pulmonary disease through regulating Nrf2 and NF-κB/NLRP3 inflammasome via directly targeting cysteine 434 of KEAP1

doi: 10.1016/j.jare.2025.07.019

Figure Lengend Snippet: Discovery of TUS as a potent therapeutic agent for COPD. (A) High-throughput screening for CXCL1 inhibitory activity of compounds from FF. RAW 264.7 macrophages were treated with 1 μg/mL LPS and 20 μM diverse compounds from FF for 24 h, and the level of CXCL1 was detected with ELISA kit. (B) Structures of sesquiterpenoids from FF. (C) The H&E or Masson’s trichrome staining images of lung tissues of mice in different groups. (D) The protein levels of iNOS, COX-2, and IL-1β in the homogenate of lung tissues of mice in different groups. (E-F) The levels of MUC5AC and SP-D in the homogenate of lung tissues of mice in different groups. (G) The percentage of neutrophil in white blood cells of mice in different groups. (H-I) The levels of TNF-α and IL-1β in BALF of mice in different groups. The results were expressed as mean ± SD (n = 4). * p < 0.05, ** p < 0.01, *** p < 0.001 treated vs CS group.

Article Snippet: The primary antibodies for KEAP1 (10503-2-AP, 1:5000), Nrf2 (16396-1-AP, 1:2000), glutamate-cystine ligase, modifier subunit (GCLM, 14241-1-AP, 1:2000), cyclooxygenase-2 (COX-2, 27308-1-AP, 1:1000), IL-1β (16806-1-AP, 1:500), β-actin (20536-1-AP, 1:30000), inducible Nitric Oxide Synthase (iNOS, 22226-1-AP, 1:2000), HA (51064-2-AP, 1:2000), inhibitor of NF-κB (IκB, 10268-1-AP, 1:2000), Caspase-1 (22915-1-AP, 1:2000), NLRP3 (27458-1-AP, 1:2000), α-tubulin (11224-1-AP, 1:10000), and the secondary antibodies HRP-conjugated goat anti-mouse IgG (66031-1-Ig, 1:10000) and HRP-conjugated goat anti-rabbit IgG (66031-2-Ig, 1:10000) were purchased from Proteintech Group (Wuhan, China).

Techniques: High Throughput Screening Assay, Activity Assay, Enzyme-linked Immunosorbent Assay, Staining

Journal: Journal of Advanced Research

Article Title: Tussilagone attenuated cigarette smoke-induced chronic obstructive pulmonary disease through regulating Nrf2 and NF-κB/NLRP3 inflammasome via directly targeting cysteine 434 of KEAP1

doi: 10.1016/j.jare.2025.07.019

Figure Lengend Snippet: TUS inhibited NF-κB by activating Nrf2. (A) The protein levels of p -NF-κB, IκB, iNOS, and COX-2 in the RAW 264.7 cells treated with 1 μg/mL LPS and TUS at 2.5, 5, 10, and 20 μM. (B) The regulation of TUS on p -NF-κB levels in the cytoplasm and nucleus. RAW 264.7 cells treated with 1 μg/mL LPS and TUS at 10 and 20 μM for 8 h, and the protein level of p -NF-κB in the cytoplasm and nucleus were detected using Western blot. (C) Immunofluorescence indicated that TUS inhibited p -NF-κB translocation into the nucleus. RAW 264.7 cells were treated with 1 μg/mL LPS and TUS at 10 and 20 μM for 8 h, followed by incubation with p -NF-κB primary antibody and Alex 594 secondary antibody. The immunofluorescence of p -NF-κB was observed and imaged. (D-E) The regulatory effects of TUS on protein levels of p -NF-κB, IκB, iNOS, and COX-2 in the WT and Nrf2-silenced RAW 264.7 cells. WT and Nrf2-silenced RAW 264.7 cells were treated with 10 μM TUS, and the protein levels of p -NF-κB, iNOS, and COX-2 were detected using Western blot. (F) The regulatory effects of TUS on protein levels of iNOS and COX-2 in the WT or C434A mutant RAW 264.7 cells. WT and C434A mutant RAW 264.7 cells were treated with 10 μM TUS, and the protein levels of p -NF-κB, iNOS, and COX-2 were detected using Western blot. The results were expressed as mean ± SD (n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 treated vs LPS group.

Article Snippet: The primary antibodies for KEAP1 (10503-2-AP, 1:5000), Nrf2 (16396-1-AP, 1:2000), glutamate-cystine ligase, modifier subunit (GCLM, 14241-1-AP, 1:2000), cyclooxygenase-2 (COX-2, 27308-1-AP, 1:1000), IL-1β (16806-1-AP, 1:500), β-actin (20536-1-AP, 1:30000), inducible Nitric Oxide Synthase (iNOS, 22226-1-AP, 1:2000), HA (51064-2-AP, 1:2000), inhibitor of NF-κB (IκB, 10268-1-AP, 1:2000), Caspase-1 (22915-1-AP, 1:2000), NLRP3 (27458-1-AP, 1:2000), α-tubulin (11224-1-AP, 1:10000), and the secondary antibodies HRP-conjugated goat anti-mouse IgG (66031-1-Ig, 1:10000) and HRP-conjugated goat anti-rabbit IgG (66031-2-Ig, 1:10000) were purchased from Proteintech Group (Wuhan, China).

Techniques: Western Blot, Immunofluorescence, Translocation Assay, Incubation, Mutagenesis

Journal: Journal of Advanced Research

Article Title: Tussilagone attenuated cigarette smoke-induced chronic obstructive pulmonary disease through regulating Nrf2 and NF-κB/NLRP3 inflammasome via directly targeting cysteine 434 of KEAP1

doi: 10.1016/j.jare.2025.07.019

Figure Lengend Snippet: TUS inhibited lung inflammation by activating Nrf2. (A) The images of lung tissues stained by H&E in indicated groups. (B-E) The levels of TNF-α and IL-1β in BALF of different groups. (F) The protein levels of Nrf2, iNOS, p -NF-κB, and IL-1β in the homogenate of lung tissues in indicated groups. (G) The mRNA levels of NF-κB, iNOS, NLRP3, COX-2, TNF-α, and IL-1β in the homogenate of lung tissues in indicated groups. The results were expressed as mean ± SD (n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 treated vs LPS group.

Article Snippet: The primary antibodies for KEAP1 (10503-2-AP, 1:5000), Nrf2 (16396-1-AP, 1:2000), glutamate-cystine ligase, modifier subunit (GCLM, 14241-1-AP, 1:2000), cyclooxygenase-2 (COX-2, 27308-1-AP, 1:1000), IL-1β (16806-1-AP, 1:500), β-actin (20536-1-AP, 1:30000), inducible Nitric Oxide Synthase (iNOS, 22226-1-AP, 1:2000), HA (51064-2-AP, 1:2000), inhibitor of NF-κB (IκB, 10268-1-AP, 1:2000), Caspase-1 (22915-1-AP, 1:2000), NLRP3 (27458-1-AP, 1:2000), α-tubulin (11224-1-AP, 1:10000), and the secondary antibodies HRP-conjugated goat anti-mouse IgG (66031-1-Ig, 1:10000) and HRP-conjugated goat anti-rabbit IgG (66031-2-Ig, 1:10000) were purchased from Proteintech Group (Wuhan, China).

Techniques: Staining

Journal: Materials Today Bio

Article Title: Ultrasound-activated piezoelectric Silk-PVDF hydrogel reprograms the osteoimmune microenvironment via NRF2 signaling for accelerated bone regeneration

doi: 10.1016/j.mtbio.2026.102779

Figure Lengend Snippet: Piezoelectric hydrogel activates NRF2 to attenuate ROS and macrophages polarization for osteogenesis . (A–B) RT-qPCR results for the mRNA expression of pro-inflammatory differentiation of macrophages. (C–D) RT-qPCR results for the mRNA expression of anti-inflammatory differentiation of macrophages. (E) The relative protein expression levels of INOS, CD206. (F–G) Semi-quantitative analysis of immunoblotting results of INOS, CD206. (H) ROS staining of ADSCs. (I) Mean intensity of ROS staining. (J) The relative protein expression levels of NRF2, NQO1, GPX4. (K–M) Semi-quantitative analysis of immunoblotting results of NRF2, NQO1, GPX4. Data are presented as the mean ± SEM; n = 3; ∗significant difference between selected groups, ∗ p < 0.05, ∗∗ p < 0.01 and ∗∗∗ p < 0.001.

Article Snippet: Cells were fixed with 4 % paraformaldehyde (Servicebio, #G1101) for 15 min at 25 °C, permeabilized with 0.3 % Triton X-100 in PBS for 15 min, and blocked with 5 % BSA (Sigma, #A7906) containing 10 % normal goat serum (Servicebio, #G5009) for 1 h. Primary antibodies were diluted in antibody diluent (Servicebio, #G1212) and incubated overnight at 4 °C: • Osteopontin (OPN): Rabbit monoclonal (Proteintech, #22952-1-AP), 1:500 • Osteocalcin (OCN): Rabbit polyclonal (Proteintech, #20277-1-AP), 1:500 • iNOS: Mouse anti-iNOS (Proteintech, #22226-1-AP), 1:500 • CD206: Rabbit anti-CD206 (Proteintech, #18704-1-AP), 1:500 After three PBS washes, species-matched secondary antibodies were applied for 1 h at 25 °C in the dark: • Alexa Fluor 488: Goat anti-rabbit IgG (Servicebio, #GB25303), 1:500 • Alexa Fluor 488: Goat anti-mouse IgG (Servicebio, #GB25301), 1:500 • Cy3: Goat anti-rabbit IgG (Servicebio, # GB21303), 1:500 Nuclei were counterstained with DAPI (Servicebio, #G1407).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Staining