anti inos nos2  (Novus Biologicals)

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: 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

Journal: The FASEB Journal

Article Title: ZXDB Drives Macrophage Inflammatory Programming in Sepsis‐Induced Acute Kidney Injury by Recruiting EIF4A3 to Enhance ACACA Translation

doi: 10.1096/fj.202502962RR

Figure Lengend Snippet: ZXDB is required for pro‐inflammatory macrophage activation and metabolic reprogramming. (A, B) Flow cytometric analysis of M1‐like (CD86 + INOS + ), M2a‐like (CD206 + ARG1 + ), and M2b‐like (CD86 + IL‐10 + ) surface markers on RAW264.7 (A) and THP‐1 (B) macrophages following stimulation with LPS (100 ng/mL) for 6 h, with or without Zxdb knockdown (shZxdb). Representative plots and quantification are shown. (C, D) Western blot analysis of key M1‐like (iNOS, CD40, CD86, CD80) and M2‐like (CD206, CD163, Arg1) protein markers in RAW264.7 (C) and THP‐1 (D) cells under the same conditions. GAPDH served as the loading control. (E, F) ELISA quantification of pro‐inflammatory (TNF‐α, IFN‐γ, IL‐1β) and anti‐inflammatory (IL‐10, IL‐4, IL‐13) cytokines secreted into the supernatant of RAW264.7 (E) and THP‐1 (F) cells. (G‐J) Assessment of metabolic state via relative lactate production (G, H) and intracellular ATP levels (I, J) in RAW264.7 and THP‐1 cells. p < 0.05, * p < 0.01, ** p < 0.001.

Article Snippet: Membranes were then incubated overnight at 4°C with primary antibodies against: ZXDB (A303‐656A; Invitrogen, 1:1000), ACACA (21923‐1‐AP; Proteintech, RRID:AB_11042445, 1:1000), EIF4A3 (17 504‐1‐AP; Proteintech, RRID:AB_2097393, 1:1000), iNOS (22226‐1‐AP; Proteintech, RRID:AB_2879038, 1:1000), CD86 (13395‐1‐AP; Proteintech–, RRID:AB_2074882, 1:1000), ARG1 (16001‐1‐AP; Proteintech–, RRID:AB_2289842, 1:1000), CD206 (18704‐1‐AP; Proteintech, RRID:AB_10597232, 1:1000), GAPDH (60004‐1‐Ig; Proteintech, RRID:AB_2107436, 1:10000).

Techniques: Activation Assay, Knockdown, Western Blot, Control, Enzyme-linked Immunosorbent Assay

Journal: The FASEB Journal

Article Title: ZXDB Drives Macrophage Inflammatory Programming in Sepsis‐Induced Acute Kidney Injury by Recruiting EIF4A3 to Enhance ACACA Translation

doi: 10.1096/fj.202502962RR

Figure Lengend Snippet: ACACA mediates the pro‐inflammatory action of ZXDB on macrophages. (A) Overlap of ZXDB‐regulated genes/proteins and ACACA‐interacting proteins. (B) Protein–protein interaction network showing ACACA's association with the ZXDB network. (C) ACACA mRNA expression in THP‐1 cells, treated with LPS (100 ng/mL) +/− shZxdb for 6 h. (D) ACACA protein expression in THP‐1 cells, treated as in (C). (E–H) Cell proliferation (E, G) and apoptosis (F, H) in RAW264.7 (E, F) and THP‐1 cells (G, H) subjected to rescue experiments (LPS +/− shZxdb +/− Acaca overexpression). (I‐J) M1‐like (CD86 + INOS + ), M2a‐like (CD206 + ARG1 + ), and M2b‐like (CD86 + IL‐10 + ) macrophage populations in RAW264.7 (I) and THP‐1 cells (J) from the rescue experiments. (K‐L) Cytokine secretion (TNF‐α, IFN‐γ, IL‐1β, IL‐10, IL‐4, IL‐13) in RAW264.7 (K) and THP‐1 cell supernatants from the rescue experiments. (M‐P) Relative ATP levels (M, N) and lactate production (O, P) in RAW264.7 (M, O) and THP‐1 cells (N, P) from the rescue experiments. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Membranes were then incubated overnight at 4°C with primary antibodies against: ZXDB (A303‐656A; Invitrogen, 1:1000), ACACA (21923‐1‐AP; Proteintech, RRID:AB_11042445, 1:1000), EIF4A3 (17 504‐1‐AP; Proteintech, RRID:AB_2097393, 1:1000), iNOS (22226‐1‐AP; Proteintech, RRID:AB_2879038, 1:1000), CD86 (13395‐1‐AP; Proteintech–, RRID:AB_2074882, 1:1000), ARG1 (16001‐1‐AP; Proteintech–, RRID:AB_2289842, 1:1000), CD206 (18704‐1‐AP; Proteintech, RRID:AB_10597232, 1:1000), GAPDH (60004‐1‐Ig; Proteintech, RRID:AB_2107436, 1:10000).

Techniques: Expressing, Over Expression

Journal: The FASEB Journal

Article Title: ZXDB Drives Macrophage Inflammatory Programming in Sepsis‐Induced Acute Kidney Injury by Recruiting EIF4A3 to Enhance ACACA Translation

doi: 10.1096/fj.202502962RR

Figure Lengend Snippet: The ZXDB‐EIF4A3 Interaction Is required for Its Pro‐inflammatory Functions. (A) Co‐IP of HA‐EIF4A3 with Flag‐ZXDB or ZXDB‐MUT in THP‐1 cells. (B) ACACA protein expression in THP‐1 cells transfected with the indicated plasmids and treated with LPS for 6 h. (C) Polysome profiling of ACACA mRNA in THP‐1 cells with Flag‐ZXDB or ZXDB‐MUT, stimulated with LPS. (D) RIP‐qPCR analysis of ACACA mRNA associated with ZXDB or ZXDB‐MUT. (E‐F) Cell proliferation (E) and apoptosis (F) in THP‐1 cells transfected and treated with LPS for 6 h. (G) M1‐like (CD86 + INOS + ), M2a‐like (CD206 + ARG1 + ) and M2b‐like (CD86 + IL‐10 + ) populations in THP‐1 cells. (H) M1‐like (iNOS, CD40, CD86, CD80) and M2‐like (CD206, CD163, ARG1) protein expression in THP‐1 cells. (I) Cytokine secretion (IFN‐γ, TNF‐α, IL‐6, IL‐1β, IL‐4, IL‐13, IL‐10) in THP‐1 cell supernatants. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Membranes were then incubated overnight at 4°C with primary antibodies against: ZXDB (A303‐656A; Invitrogen, 1:1000), ACACA (21923‐1‐AP; Proteintech, RRID:AB_11042445, 1:1000), EIF4A3 (17 504‐1‐AP; Proteintech, RRID:AB_2097393, 1:1000), iNOS (22226‐1‐AP; Proteintech, RRID:AB_2879038, 1:1000), CD86 (13395‐1‐AP; Proteintech–, RRID:AB_2074882, 1:1000), ARG1 (16001‐1‐AP; Proteintech–, RRID:AB_2289842, 1:1000), CD206 (18704‐1‐AP; Proteintech, RRID:AB_10597232, 1:1000), GAPDH (60004‐1‐Ig; Proteintech, RRID:AB_2107436, 1:10000).

Techniques: Co-Immunoprecipitation Assay, Expressing, Transfection

Journal: The FASEB Journal

Article Title: ZXDB Drives Macrophage Inflammatory Programming in Sepsis‐Induced Acute Kidney Injury by Recruiting EIF4A3 to Enhance ACACA Translation

doi: 10.1096/fj.202502962RR

Figure Lengend Snippet: Schematic diagram of the proposed mechanism by which ZXDB promotes M1‐like macrophage polarization and exacerbates SI‐AKI. In macrophage, ZXDB interacts with EIF4A3, promoting the translation of the ACACA gene. The resulting increase in ACACA protein expression enhances glycolysis and lactate production. This metabolic reprogramming shifts macrophage polarization toward M1‐like macrophage activation (characterized by increased Cd86, Cd80, Cd40, and iNOS) and away from an anti‐inflammatory M2‐like phenotype (characterized by Cd206, Cd163, and Arg1). The dominance of M1‐like macrophages leads to an elevated secretion of pro‐inflammatory cytokines (TNF‐α, IFN‐γ, IL‐1β) and reduced anti‐inflammatory cytokines (IL‐10, IL‐4, IL‐13), which collectively drive the pathogenesis of acute kidney injury.

Article Snippet: Membranes were then incubated overnight at 4°C with primary antibodies against: ZXDB (A303‐656A; Invitrogen, 1:1000), ACACA (21923‐1‐AP; Proteintech, RRID:AB_11042445, 1:1000), EIF4A3 (17 504‐1‐AP; Proteintech, RRID:AB_2097393, 1:1000), iNOS (22226‐1‐AP; Proteintech, RRID:AB_2879038, 1:1000), CD86 (13395‐1‐AP; Proteintech–, RRID:AB_2074882, 1:1000), ARG1 (16001‐1‐AP; Proteintech–, RRID:AB_2289842, 1:1000), CD206 (18704‐1‐AP; Proteintech, RRID:AB_10597232, 1:1000), GAPDH (60004‐1‐Ig; Proteintech, RRID:AB_2107436, 1:10000).

Techniques: Expressing, Activation Assay