Journal: Journal of Animal Science and Biotechnology

Article Title: Dietary supplementation of benzoic acid and essential oils combination enhances intestinal resilience against LPS stimulation in weaned piglets

doi: 10.1186/s40104-023-00958-6

Figure Lengend Snippet: Primer sequences used in real-time PCR

Article Snippet: After that, the PVDF membranes was blocked with corresponding primary antibody at 4 °C for 15 h. Antibodies against Claudin-1 (1:1,000, ab129119, Abcam, Cambridge, UK), ZO-1 (1:3,000, 21773-1-AP, Proteintech, Wuhan, China), Occludin (1:2,000, 27260-1-AP, Proteintech), P-JNK (1:1,000, 4688S, Cell Signaling Technology, Boston, USA), JNK (1:1,000, 9252S, Cell Signaling Technology), P-ERK(1:2,000, 9101S, Cell Signaling Technology), ERK (1:2,000, 9102S, Cell Signaling Technology), P-P38 (1:2,000, 4092S, Cell Signaling Technology), P38 (1:3,000, 66234-1-Ig, Proteintech), P-NF-κB (1:1,000, 3033S, Cell Signaling Technology), NF-κB (1:1,000, 10745-1-AP, Proteintech), P-Nrf2 (1:1,000, 381559, ZenBio, Chengdu, China), Nrf2 (1:1,000, 16396-1-AP, Proteintech), Keap1 (1:2,000, 16396-1-AP, Proteintech) and β-actin (1:2,000, bs-0061R, Bioss, Beijing, China).

Techniques:

Journal: Journal of Animal Science and Biotechnology

Article Title: Dietary supplementation of benzoic acid and essential oils combination enhances intestinal resilience against LPS stimulation in weaned piglets

doi: 10.1186/s40104-023-00958-6

Figure Lengend Snippet: Effects of dietary BAO supplementation on protein and mRNA expressions of MAPK/NF-κB inflammatory pathway in jejunum of weaned pigs challenged with LPS. A – E The protein expression levels of P-ERK/ERK, P-JNK/JNK, P-P38/P38 and P-NF-κB/NF-κB. F – I The mRNA expression levels of ERK , JNK , P38 and NF-κB . Dates are presented as means ± SEM ( n = 3 for protein expression; n = 5 for mRNA expression). * P < 0.05; ** P < 0.01. CON, corn-soybean meal basal diets group treated with saline; LPS, corn-soybean meal basal diets group treated with lipopolysaccharide; BAO + LPS, corn-soybean meal basal diets + 5 kg/t benzoic acid + 500 g/t essential oils group treated with lipopolysaccharide

Article Snippet: After that, the PVDF membranes was blocked with corresponding primary antibody at 4 °C for 15 h. Antibodies against Claudin-1 (1:1,000, ab129119, Abcam, Cambridge, UK), ZO-1 (1:3,000, 21773-1-AP, Proteintech, Wuhan, China), Occludin (1:2,000, 27260-1-AP, Proteintech), P-JNK (1:1,000, 4688S, Cell Signaling Technology, Boston, USA), JNK (1:1,000, 9252S, Cell Signaling Technology), P-ERK(1:2,000, 9101S, Cell Signaling Technology), ERK (1:2,000, 9102S, Cell Signaling Technology), P-P38 (1:2,000, 4092S, Cell Signaling Technology), P38 (1:3,000, 66234-1-Ig, Proteintech), P-NF-κB (1:1,000, 3033S, Cell Signaling Technology), NF-κB (1:1,000, 10745-1-AP, Proteintech), P-Nrf2 (1:1,000, 381559, ZenBio, Chengdu, China), Nrf2 (1:1,000, 16396-1-AP, Proteintech), Keap1 (1:2,000, 16396-1-AP, Proteintech) and β-actin (1:2,000, bs-0061R, Bioss, Beijing, China).

Techniques: Expressing, Saline

Journal: Advanced Science

Article Title: Engineered Microparticles for Treatment of Murine Brain Metastasis by Reprograming Tumor Microenvironment and Inhibiting MAPK Pathway

doi: 10.1002/advs.202206212

Figure Lengend Snippet: Cytokine detection after treatment and molecular mechanisms of macrophage reprogramming induced by P5091@RMPs‐R4F. A) Cytokine concentrations in serum from the indicated treatment groups as measured by flow cytometry ( n = 6 mice). B) Heat map illustrating the differentially expressed M1‐and M2‐related genes in TAMs in the P5091@RMPs‐R4F group and the Control group based on RNA sequencing results. C) KEGG analysis identifying the 17 most enriched pathways based on the differentially expressed genes of the two groups. D) Volcano plots of the differentially expressed genes. Red dots show significantly up‐regulated genes, and green dots show significantly down‐regulated genes. E) Western blotting of p‐JNK, p‐ERK, p‐p38, and GAPDH in IL‐4/13‐BMDM M2 cells treated with P5091@RMPs‐R4F at the indicated time points. Statistical analysis was performed using one‐way ANOVA with Tukey's multiple comparison test for (A). Data are presented as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and ns: not significant.

Article Snippet: SR‐B1 antibody (Catalog number: 21277‐1‐AP), TSG101 antibody (Catalog number: 67381‐1‐Ig), CD9 antibody (Catalog number: 20597‐1‐AP), Phospho‐JNK (Tyr185) Recombinant antibody (Catalog number: 80024‐1‐RR), Phospho‐ERK1/2 (Thr202/Tyr204) Polyclonal antibody (Catalog number: 28733‐1‐AP), and Phospho‐p38 MAPK (Thr180/Tyr182) Polyclonal antibody (Catalog number: 28796‐1‐AP) were purchased from Proteintech.

Techniques: Flow Cytometry, Control, RNA Sequencing, Western Blot, Comparison

Journal: Cells

Article Title: Functional and Phenotypic Characterization of Siglec-6 on Human Mast Cells

doi: 10.3390/cells11071138

Figure Lengend Snippet: Co-crosslinking of Siglec-6 and FcεRIα on MCs enhances inhibitory activity. ( A , B ) CD34+ cell-derived MCs were incubated with the indicated concentration of anti-FcεRIα, as well as anti-Siglec-6 or isotype control mAb. Antibodies were crosslinked using secondary anti-mouse IgG antibody, and cells were incubated at 37 °C for 20 min to permit cell stimulation. The percentage of CD63+ ( A ) or LAMP-1+ ( B ) MCs was then determined by flow cytometry. ( C – G ) CD34+ cell-derived MCs were incubated with 150 ng/mL anti-FcεRIα and either anti-Siglec-6 or isotype control mAb, or were not incubated with mAbs (No Stim). Expression of CD63 ( C ) was determined by flow cytometry, and tryptase release ( D ) was measured colorimetrically in cell-free supernatant after 20 min of stimulation. MIP-1β ( E ), IL-8 ( F ), and IL-1β ( G ) were detected in cell-free supernatant after overnight stimulation. Levels were normalized to those measured in the stimulated samples incubated with the isotype control antibody ( D – G ). ( H – J ) CD34+ cell-derived MCs were incubated in the presence or absence (No Stim) of 150 ng/mL anti-FcεRIα with either anti-Siglec-6 or isotype control mAb. Antibodies were crosslinked using secondary anti-mouse IgG antibody, and cells were incubated at 37 °C for 15 min in complete medium to permit cell stimulation. Following stimulation, cells were fixed and stained for surface CD63 ( H ), intracellular phospho-ERK1/2 ( I ), or intracellular phospho-p38 ( J ). Data are representative ( A , B ), represent the means and standard deviations of four ( C – E ) or three ( F , G ) independent mast cell cultures and experiments, or represent the individual values, means, and standard deviations of two ( H – J : No Stim) or four ( H – J : Anti-FcεRIα) replicates. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 by one-way ANOVA with Tukey test to correct for multiple comparisons ( C – G ). **** p < 0.0001 by two-way ANOVA with Šidák correction for multiple comparisons ( H – J ).

Article Snippet: The cells were then washed in FACS buffer, permeabilized by resuspending in cold (−20 °C) methanol, added slowly while shaking, then incubated on ice for 30 min and stained with anti-phospho-ERK1/2 (Thr202/Tyr204, clone 197G2, Cell Signaling Technology, Danvers, MA, USA) or anti-phospho-p38 (Thr180/Tyr182, clone 28B10, Cell Signaling Technology).

Techniques: Activity Assay, Derivative Assay, Incubation, Concentration Assay, Control, Cell Stimulation, Flow Cytometry, Expressing, Staining