Journal: Frontiers in Nutrition

Article Title: High intake of n-6 polyunsaturated fatty acid exacerbates non-alcoholic steatohepatitis by the involvement of multiple metabolic pathways

doi: 10.3389/fnut.2025.1562509

Figure Lengend Snippet: Effects of n-6 PUFA on liver macrophage phenotype in rats with NASH induced by a choline-deficient diet. (A) M1-type Kupffer cells (KCs) identified by double staining: red arrows show CD11c-positive cells, green arrows show CD68-positive cells, and yellow arrows highlight CD11c and CD68 double-positive M1-type KCs (Scale bar – 50 μM). (B) M2-type KCs identified similarly, with red arrows indicating CD163-positive cells, green arrows showing CD68-positive cells, and yellow arrows marking CD163 and CD68 double-positive M2-type KCs (Scale bar – 50 μM). For (A,B) (see ) for full-size photomicrographs. (C) M1/M2 phenotype ratio (unitless), calculated as the proportion of CD68 + CD11c + to CD68 + CD163 + cells. (D) Relative PPAR-γ2 mRNA expression (fold change normalized to GAPDH) in the liver, which is linked to macrophage polarization and inflammation. Data are expressed as mean ±SEM; n = 6/group.

Article Snippet: BCA protein quantitation kit (Boster Bio, Cat # AR0146), RIPA lysis buffer (Boster Bio, Cat # 0105), protease inhibitor cocktails (Boster, Bio Cat # AR1182), phosphatase inhibitor (Boster Bio, Cat # AR1183), color pre-dyed protein marker (Boster Bio, Cat # AR1113), Western-specific primary and secondary antibody diluent (Boster Bio, Cat # AR1017), wash buffer TBS-T (Boster Bio, Cat # AR0195-10), ECL chemiluminescent reagent (Boster Bio, Cat # AR1196), BSA TBS buffer system blocking solution (Boster Bio, Cat # AR0189), NF-κB antibody (Boster Bio, Cat # A01228-1), GAPDH antibody (Boster Bio, Cat # M00227), HRP-conjugated goat anti-rabbit IgG (Boster Bio, Cat # BA1054), CD163 antibody (Boster Bio, Cat # A00812-2), CD11c antibody (Boster Bio, Cat # A00357-3), CD68 antibody (Boster Bio, Cat # BA3638), fluorescent (DyLight 488) labeled goat anti-rabbit IgG (Boster Bio, Cat # BA1127), fluorescent (DyLight 594) labeled goat anti-rabbit IgG (Boster Bio, Cat # BA1142), rat TNF-α ELISA kit (Jiangsu Enzyme Immunoassay Co., Ltd., Cat # MM-0180R1), rat IL-1β ELISA kit (Boster Bio, Cat # EK0393), rat IL-4 ELISA kit (Jiangsu Enzyme Immunoassay Co., Ltd., Cat # MM-0191R1), rat IL-10 ELISA kit (Boster Bio, Cat # EK0418), 25 VD3 assay kit (Roche Diagnostics, Cat # 07028148190), TAOC assay kit (Nanjing Jiancheng, Cat # A015-3-1), MDA assay kit (Nanjing Jiancheng, Cat # A003-1-2), and free fatty acid kit (Kunchuang Biotechnology, Xian, China, Cat # SK125-2).

Techniques: Double Staining, Expressing

Journal: Scientific Reports

Article Title: Immuno-targeting the multifunctional CD38 using nanobody

doi: 10.1038/srep27055

Figure Lengend Snippet: ( a ) The structure of 1053-PE38. ( b ) SDS-PAGE analysis of pure 1053-PE38 and GFPNb-PE38. The cytotoxicity of 1053-PE38 ( c ) and GFPNb-PE38 ( d ), as a control, on MM cell lines, with or without RA pre-treatment measured by WST-1 assay and analyzed by Graphpad. The experiments were performed four times.

Article Snippet: The sequence encoding hinge region of human CD8 and the truncated Pseudomonas exotoxin A (PE38) was amplified from the plasmid anti-CD11c IMMUNOTOXIN from Addgene (#22850) and subcloned into the above plasmid pET28a-1053 by Hind III and Not I.

Techniques: SDS Page, Control, WST-1 Assay

Journal: Scientific Reports

Article Title: Immuno-targeting the multifunctional CD38 using nanobody

doi: 10.1038/srep27055

Figure Lengend Snippet: ( a ) CD38 expression in five different MM patient samples and PWBCs were analyzed by 1053-EGFP staining method, together with LP-1 and CD38-KO as relative expression controls. ( b ) The cytotoxicity of 1053-PE38 after 3-day treatment was analyzed by calcein staining followed by FACS. Three repeats were performed on each batch of samples.

Article Snippet: The sequence encoding hinge region of human CD8 and the truncated Pseudomonas exotoxin A (PE38) was amplified from the plasmid anti-CD11c IMMUNOTOXIN from Addgene (#22850) and subcloned into the above plasmid pET28a-1053 by Hind III and Not I.

Techniques: Expressing, Staining

Journal: Iranian Journal of Basic Medical Sciences

Article Title: Expressions of CD11a, CD11b, and CD11c integrin proteins in rats with myocardial hypertrophy

doi:

Figure Lengend Snippet: Oligonucleotide primers of CD11, CD11b, CD11c, and β-actin

Article Snippet: Primary antibodies of CD11a, CD11b, and CD11c (1:100, Wuhan Boster Company) were incubated overnight at 4°C and then incubated at 37°C for 30 min in IgG/Biotin IgG and SABC liquid.

Techniques:

Journal: Iranian Journal of Basic Medical Sciences

Article Title: Expressions of CD11a, CD11b, and CD11c integrin proteins in rats with myocardial hypertrophy

doi:

Figure Lengend Snippet: RT-PCR products of CD11a, CD11b, and CD11c A: RT-PCR products of CD11a M: Marker 1-2: experimental group; 3-4: control group B: RT-PCR products of CD11b M: Marker 1-2: control group; 3-4: experimental group C: RT-PCR products of CD11c M: Marker 1-2: experimental group; 3-4: control group

Article Snippet: Primary antibodies of CD11a, CD11b, and CD11c (1:100, Wuhan Boster Company) were incubated overnight at 4°C and then incubated at 37°C for 30 min in IgG/Biotin IgG and SABC liquid.

Techniques: Reverse Transcription Polymerase Chain Reaction, Marker, Control

Journal: Iranian Journal of Basic Medical Sciences

Article Title: Expressions of CD11a, CD11b, and CD11c integrin proteins in rats with myocardial hypertrophy

doi:

Figure Lengend Snippet: Comparison of the average area and brightness of CD11a, CD11b, and CD11c expressions in the myocardium ( x ¯ ±s)

Article Snippet: Primary antibodies of CD11a, CD11b, and CD11c (1:100, Wuhan Boster Company) were incubated overnight at 4°C and then incubated at 37°C for 30 min in IgG/Biotin IgG and SABC liquid.

Techniques: Comparison, Control

Journal: Iranian Journal of Basic Medical Sciences

Article Title: Expressions of CD11a, CD11b, and CD11c integrin proteins in rats with myocardial hypertrophy

doi:

Figure Lengend Snippet: Expressions of CD11a, CD11b, and CD11c in the myocardial tissue at mRNA levels A: CD11a of control group B: CD11b of control group C: CD11c of control group D: CD11a of experimental group E: CD11b of experimental group F: CD11c of experimental group Arrows was used to indicate the positive expression of integrins in experimental group.

Article Snippet: Primary antibodies of CD11a, CD11b, and CD11c (1:100, Wuhan Boster Company) were incubated overnight at 4°C and then incubated at 37°C for 30 min in IgG/Biotin IgG and SABC liquid.

Techniques: Control, Expressing

Journal: Journal of Nanobiotechnology

Article Title: Lactiplantibacillus plantarum -derived extracellular vesicles alleviate acute lung injury by inhibiting ferroptosis of macrophages

doi: 10.1186/s12951-025-03405-y

Figure Lengend Snippet: LpEVs alleviated lung inflammation and injury in mice. A . Schematic depiction of the experiment. B . Representative images showing histopathological alterations in lungs of different treatment groups. ICR mice ( n = 5) were sitimulated with LPS (2.5 mg/kg, intratracheally) for 3 h, then administered PBS or LpEVs (100 µg) via the tail vein injection method. Mice were euthanized 24 h post-treatment. Scale bar = 200 μm. C . Representative F4/80 and MPO IHC staining images in different lungs. Scale bars represent 50 μm. D . Protein concentration in BALF. E . Cell counts in BALF. F . The concentration of cytokines/chemokines in the BALF was assessed using ELISA. G . Flow cytometry analysis of neutrophils (CD11b + Ly6G + ) in the BALF. H . Flow cytometry analysis of macrophages (CD11b + F4/80 + ) in the BALF. I . Flow cytometry analysis of alveolar macrophages (CD11c + Siglec F + ) in the BALF

Article Snippet: The population of alveolar macrophages (anti-CD11c, anti-Siglec F) (E-AB-F0991E, Elabscience, Wuhan, China) (12-1702-82, Thermo Fisher, USA), macrophages (anti-F4/80, anti-CD11b) (E-AB-F0995D, E-AB-F1081E, Elabscience, Wuhan, China), and neutrophils (anti-CD11b, anti-Ly6G) (E-AB-F1081E, E-AB-F1108C, Wuhan, China) were assessed using flow cytometry with specific antibodies.

Techniques: Injection, Immunohistochemistry, Protein Concentration, Concentration Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: Journal of Nanobiotechnology

Article Title: Lactiplantibacillus plantarum -derived extracellular vesicles alleviate acute lung injury by inhibiting ferroptosis of macrophages

doi: 10.1186/s12951-025-03405-y

Figure Lengend Snippet: LpEVs treatment inhibited LPS-induced ferroptosis in vivo. A . The levels of reactive oxygen species (ROS) in lung BALF cells were evaluated using DCFH-DA. The left panel displays a flow cytometry diagram, while the right panel shows the mean fluorescence intensity of ROS. B . Relative MDA content in lungs. C . Relative Fe 2+ content in lungs. D . Relative GSH content in lungs. E . Relative mRNA expression of ferroptotic-related genes were analyzed by qRT-PCR. F . The expression levels of ferroptotic-related proteins in lungs were analyzed by western blotting. ACTIN is adopted as the loading control. G . Representative IHC staining images for GPX4, SLC7A11 and ACSL4 were obtained from different lung samples. Scale bars represent 50 μm. H . The statistical graph of the expression of GPX4, SLC7A11 and ACSL4 in IHC staining. I . Representative IHC staining images for macrophages M1 marker CD86 and M1 marker CD206 in lungs. Scale bars represent 50 μm. J . The statistical graph of the expression of CD86 and CD206 in IHC staining. Quantification of proteins expresion levels in IHC staining were determined by image J software. The values for the LPS-PBS and LPS-EV were expressed as percentages of the values for the Sham group. * P < 0.05, ** P < 0.01, *** P < 0.001, *** P < 0.0001 and ns, no significant difference

Article Snippet: The population of alveolar macrophages (anti-CD11c, anti-Siglec F) (E-AB-F0991E, Elabscience, Wuhan, China) (12-1702-82, Thermo Fisher, USA), macrophages (anti-F4/80, anti-CD11b) (E-AB-F0995D, E-AB-F1081E, Elabscience, Wuhan, China), and neutrophils (anti-CD11b, anti-Ly6G) (E-AB-F1081E, E-AB-F1108C, Wuhan, China) were assessed using flow cytometry with specific antibodies.

Techniques: In Vivo, Flow Cytometry, Fluorescence, Expressing, Quantitative RT-PCR, Western Blot, Control, Immunohistochemistry, Marker, Software

Journal: Journal of Nanobiotechnology

Article Title: Lactiplantibacillus plantarum -derived extracellular vesicles alleviate acute lung injury by inhibiting ferroptosis of macrophages

doi: 10.1186/s12951-025-03405-y

Figure Lengend Snippet: LpEVs facilitated the transition of macrophages towards an anti-inflammatory phenotype. A . Confocal laser scanning microscopy images displayed the presence of Dio-labeled LpEVs in MH-S cells. Green: Dio labeled LpEVs; Blue: Hoechst. Scale bar = 20 μm. B . The Dio-positive MH-S cells were determined by flow cytometry. C . Relative mRNA expression levels of inflammatory factors were assessed by qRT-PCR. M1 proinflammatory genes: Il1β , Il6 , Tnfα , and iNos , M2 anti-inflammatory genes: Arg-1 , and Cd206 . The M1 macrophages were induced by LPS treatment, and M2 macrophages were induced by IL-4 treatment. D . The levels of pro-inflammatory and anti-inflammatory cytokines in the medium supernatants were quantified using ELISA. E . The expression of CD86 (an M1 marker) on MH-S cells 12 h after exposure to LPS was assessed using flow cytometry, with representative histograms and average relative mean fluorescence intensity (MFI) provided. F . Flow cytometry was used to detect the expression of CD206, an M2 marker, on MH-S cells 12 h after exposure to IL4

Article Snippet: The population of alveolar macrophages (anti-CD11c, anti-Siglec F) (E-AB-F0991E, Elabscience, Wuhan, China) (12-1702-82, Thermo Fisher, USA), macrophages (anti-F4/80, anti-CD11b) (E-AB-F0995D, E-AB-F1081E, Elabscience, Wuhan, China), and neutrophils (anti-CD11b, anti-Ly6G) (E-AB-F1081E, E-AB-F1108C, Wuhan, China) were assessed using flow cytometry with specific antibodies.

Techniques: Confocal Laser Scanning Microscopy, Labeling, Flow Cytometry, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Marker, Fluorescence

Journal: Journal of Nanobiotechnology

Article Title: Lactiplantibacillus plantarum -derived extracellular vesicles alleviate acute lung injury by inhibiting ferroptosis of macrophages

doi: 10.1186/s12951-025-03405-y

Figure Lengend Snippet: LpEVs attenuated ferroptosis in macrophages. A . The ROS level in MH-S was assessed by flow cytometry. B . Relative MDA content in MH-S. C . Relative GSH content in MH-S. D . The intercellular Fe 2+ was detected by FerroOrange (a specific probe of Fe 2+ ). Scale bar = 100 μm. E . The expression levels of ferroptotic-related proteins and NRF2-related proteins in MH-S were detected by western blot. ACTIN is adopted as the loading control. F . SLC7A11, GPX4 and ACSL4 immunofluorescent staining results of differently treated MH-S. Scale bar = 20 μm. G. NRF2 and HO-1 immunofluorescent staining results of differently treated MH-S. Scale bar = 20 μm

Article Snippet: The population of alveolar macrophages (anti-CD11c, anti-Siglec F) (E-AB-F0991E, Elabscience, Wuhan, China) (12-1702-82, Thermo Fisher, USA), macrophages (anti-F4/80, anti-CD11b) (E-AB-F0995D, E-AB-F1081E, Elabscience, Wuhan, China), and neutrophils (anti-CD11b, anti-Ly6G) (E-AB-F1081E, E-AB-F1108C, Wuhan, China) were assessed using flow cytometry with specific antibodies.

Techniques: Flow Cytometry, Expressing, Western Blot, Control, Staining

Journal: Journal of Nanobiotechnology

Article Title: Lactiplantibacillus plantarum -derived extracellular vesicles alleviate acute lung injury by inhibiting ferroptosis of macrophages

doi: 10.1186/s12951-025-03405-y

Figure Lengend Snippet: LpEVs regulated ferroptosis in macrophages via delivery of cbn-let-7. A . miRNA microarray analyze the miRNA expression profile of LpEVs. B . The 4 miRNAs (cel-mir-4937, cbn-let-7, ngi-miR-92a and gsa-mir-9394b) expression level in LpEVs. C . cbn-let-7 expression in PBS and LpEVs treated MH-S cells. D , E . Structure and luciferase result of dual luciferase reporter gene. F . The relative expression of cbn-let-7 in NC, cbn-let-7 mimic and inhibitor treated MH-S cells. G . The protein expression of of ACSL4 and GPX4 in NC, cbn-let-7 mimic and inhibitor treated MH-S cells. H . The expression of cbn-let-7 in different treated cells. I . The protein expression of of ACSL4 and GPX4 in different treated cells. ACTIN is adopted as the loading control. J . The intercellular Fe 2+ in different treated cells was detected by FerroOrange (a specific probe of Fe 2+ )

Article Snippet: The population of alveolar macrophages (anti-CD11c, anti-Siglec F) (E-AB-F0991E, Elabscience, Wuhan, China) (12-1702-82, Thermo Fisher, USA), macrophages (anti-F4/80, anti-CD11b) (E-AB-F0995D, E-AB-F1081E, Elabscience, Wuhan, China), and neutrophils (anti-CD11b, anti-Ly6G) (E-AB-F1081E, E-AB-F1108C, Wuhan, China) were assessed using flow cytometry with specific antibodies.

Techniques: Microarray, Expressing, Luciferase, Control