Journal: Biomedicines

Article Title: Targeting the Glutamine Transporter SLC1A5 Enhances Sensitivity of Acute Myeloid Leukemia to MLN4924

doi: 10.3390/biomedicines14030667

Figure Lengend Snippet: Effect of MLN4924 in combination with SLC1A5 inhibitor (V9302) on AML cell proliferation and cell differentiation. ( A – C ) Evaluate the interference effect of shRNA on SLC1A5 in AML cells through RT-qPCR and Western blot. ( D , E ) The effect of SLC1A5 knockdown on the sensitivity to MLN4924 of AML cells was detected through CCK8 assay (24 h after treatment). ( F , G ) Kasumi-1 and MOLM-13 cells were treated with V9302 for 24 h and IC50 was detected through CCK8 assay. ( H – M ) CCK8 assay was used to verify the effect of MLN4924 combined with V9302 on AML cell viability. ( N – V ) CD11b, CD14, and CD16 expression on AML cell surface treated with MLN4924 and V9302 for 48 h was detected through flow cytometry. ( W ) Wright–Giemsa staining to detect morphological changes of AML cells treated with MLN4924 and V9302 for 24 h captured using an oil immersion lens (1000×). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Antibody against CD11b (Anti-Human CD11b Mouse IgG2a Recombinant Antibody, FITC-65582) was purchased from Proteintech (China).

Techniques: Cell Differentiation, shRNA, Quantitative RT-PCR, Western Blot, Knockdown, CCK-8 Assay, Expressing, Flow Cytometry, Staining

Journal: International Journal of Molecular Sciences

Article Title: Bupleuri Radix Polysaccharides Alleviate MASLD by Regulating Muribaculaceae -Derived SCFAs in the Gut–Liver Axis

doi: 10.3390/ijms27020637

Figure Lengend Snippet: Bupleuri radix polysaccharides (BRP) attenuates liver injury and metabolic dysfunction in methionine- and choline-deficient (MCD) diet-induced metabolic dysfunction-associated steatotic liver disease (MASLD) mice. ( A ) Schematic illustration of the experimental design. BRP (200 or 400 mg/kg) was administered to MCD diet-induced mice. ( B ) The body weight of mice in different groups. ( C ) Liver coefficient of mice in each group. ( D ) Hematoxylin and eosin (H&E) staining of liver tissues and ( E ) quantification of the extent of liver damage using the MASLD activity score (MAS) method. Scale bar = 100 μm. ( F ) The levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), total cholesterol (TC), triglyceride (TG), and non-esterified fatty acids (NEFA) in the serum. ( G ) The levels of TC, TG, and NEFA in the livers. Relative mRNA expression levels of ( H ) fibrosis-related genes Acta2 and Col1a1 and ( I ) inflammation-related markers Cd11b , Cd68 , Ccl2 , Tgfβ1 , and Tnfα in livers. Statistical significance: Data are presented as mean ± standard deviation (SD) (n = 6 mice per group). Statistical analysis was performed using one-way ANOVA followed by appropriate post hoc tests, as described in . # p < 0.05, ## p < 0.01, ### p < 0.001 compared between the control (CT) group and the model group. * p < 0.05, ** p < 0.01, *** p < 0.001 compared between the model group and the BRP treatment groups.

Article Snippet: Sections were then blocked with 0.2% Triton X-100 + 2.5% BSA + 10% goat serum blocking buffer at RT for 30 min. After blocking, primary antibody cocktails were applied: Villin (Proteintech Group Inc., Rosemont, IL, USA; 66096-1-Ig) + Ki67 (Proteintech Group Inc., Rosemont, IL, USA; 27309-1-AP) or CD11b (Proteintech Group Inc., Rosemont, IL, USA; 66519-1-Ig) + E-cadherin (Cell Signaling Technology, Inc., Danvers, MA, USA; 3195S).

Techniques: Staining, Activity Assay, Expressing, Standard Deviation, Control

Journal: International Journal of Molecular Sciences

Article Title: Bupleuri Radix Polysaccharides Alleviate MASLD by Regulating Muribaculaceae -Derived SCFAs in the Gut–Liver Axis

doi: 10.3390/ijms27020637

Figure Lengend Snippet: BRP alleviates colonic mucosal injury and inflammation during MASLD. ( A – D ) Representative ( A ) H&E staining and ( C ) AB-PAS staining of colon tissues, and ( B , D ) the quantification of colonic injury area was performed using Image J software, version 1.53. In H&E staining, nuclei are stained blue-purple and cytoplasm appears pink; in AB-PAS staining, acidic mucins are stained blue and neutral mucins magenta. Scale bar = 100 μm. Representative immunofluorescence co-staining images of ( E ) Villin (green) and Ki67 (red), as well as ( F ) Cd11b (green) and E-cadherin (red), in colon tissue. DAPI was used for nuclear labeling (blue), and yellow signals in merged images indicate colocalization. Scale bar = 100 μm. ( G ) Protein levels of E-cadherin and Occludin in colonic tissues. ( H , I ) Relative mRNA levels of Ecad , Ocln , Muc2 , Zo1 , Cd11b , Il1b , and Ccl2 in colon tissues. Statistical significance: Data are presented as mean ± SD (n = 6 mice per group). Statistical analysis was performed using one-way ANOVA followed by appropriate post hoc tests, as described in the . # p < 0.05, ## p < 0.01, ### p < 0.001 compared between the CT group and the model group. * p < 0.05, ** p < 0.01, *** p < 0.001 compared between the model group and the BRP treatment groups.

Article Snippet: Sections were then blocked with 0.2% Triton X-100 + 2.5% BSA + 10% goat serum blocking buffer at RT for 30 min. After blocking, primary antibody cocktails were applied: Villin (Proteintech Group Inc., Rosemont, IL, USA; 66096-1-Ig) + Ki67 (Proteintech Group Inc., Rosemont, IL, USA; 27309-1-AP) or CD11b (Proteintech Group Inc., Rosemont, IL, USA; 66519-1-Ig) + E-cadherin (Cell Signaling Technology, Inc., Danvers, MA, USA; 3195S).

Techniques: Staining, Software, Immunofluorescence, Labeling

Journal: International Journal of Molecular Sciences

Article Title: Bupleuri Radix Polysaccharides Alleviate MASLD by Regulating Muribaculaceae -Derived SCFAs in the Gut–Liver Axis

doi: 10.3390/ijms27020637

Figure Lengend Snippet: The hepatoprotective effect of BRP is suppressed by removing gut microbiota in MASLD mice. ( A ) Animal experimental flowchart. ( B ) The body weight of mice in different groups. ( C ) Liver coefficient of mice in each group. ( D ) H&E staining of liver tissues, and ( E ) quantification of the extent of liver damage using the MAS method. Scale bar = 100 μm. ( F ) The levels of AST, ALT, TC, and TG in the serum. ( G ) The levels of TG and TC in the livers. ( H , I ) Relative mRNA expression levels of Col1a1 , Acta2 , Cd11b , Cd68 , Tgfβ1 , Tnfα , and Ccl2 in livers. Statistical significance: Data are presented as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by appropriate post hoc tests, as described in the . n = 6 mice per group. # p < 0.05, ## p < 0.01, ### p < 0.001 compared between the CT group and either the CT + ABX group or the model + ABX group.

Article Snippet: Sections were then blocked with 0.2% Triton X-100 + 2.5% BSA + 10% goat serum blocking buffer at RT for 30 min. After blocking, primary antibody cocktails were applied: Villin (Proteintech Group Inc., Rosemont, IL, USA; 66096-1-Ig) + Ki67 (Proteintech Group Inc., Rosemont, IL, USA; 27309-1-AP) or CD11b (Proteintech Group Inc., Rosemont, IL, USA; 66519-1-Ig) + E-cadherin (Cell Signaling Technology, Inc., Danvers, MA, USA; 3195S).

Techniques: Staining, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Bupleuri Radix Polysaccharides Alleviate MASLD by Regulating Muribaculaceae -Derived SCFAs in the Gut–Liver Axis

doi: 10.3390/ijms27020637

Figure Lengend Snippet: P. intestinale synergistically enhances the hepatoprotective effect of BRP on MALFD mice. ( A ) Animal experimental flowchart. ( B ) The body weight of mice in different groups. ( C ) Liver coefficient of mice in each group. ( D ) H&E staining of liver tissues and ( E ) quantification of the extent of liver damage using the MAS method. Scale bar = 100 μm. ( F ) The levels of AST, ALT, TC, TG, and NEFA in the serum. ( G ) The levels of TC, TG, and NEFA in the livers. ( H , I ) Relative mRNA expression levels of Acta2 , Col1a1 , Cd11b , Cd68 , Ccl2 , Tgfβ1 , and Tnfα in livers. ( J ) The levels of SCFAs (acetic, propionic, butyric, pentanoic, and isovaleric acids) in cecum. Statistical significance: Data are presented as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by appropriate post hoc tests, as described in the . n = 6 mice per group. # p < 0.05, ## p < 0.01, ### p < 0.001 compared between the CT group and the model group. * p < 0.05, ** p < 0.01, *** p < 0.001 compared between the model group and the treatment groups.

Article Snippet: Sections were then blocked with 0.2% Triton X-100 + 2.5% BSA + 10% goat serum blocking buffer at RT for 30 min. After blocking, primary antibody cocktails were applied: Villin (Proteintech Group Inc., Rosemont, IL, USA; 66096-1-Ig) + Ki67 (Proteintech Group Inc., Rosemont, IL, USA; 27309-1-AP) or CD11b (Proteintech Group Inc., Rosemont, IL, USA; 66519-1-Ig) + E-cadherin (Cell Signaling Technology, Inc., Danvers, MA, USA; 3195S).

Techniques: Staining, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Bupleuri Radix Polysaccharides Alleviate MASLD by Regulating Muribaculaceae -Derived SCFAs in the Gut–Liver Axis

doi: 10.3390/ijms27020637

Figure Lengend Snippet: P. intestinale synergistically enhances BRP in improving colonic mucosal damage in MALFD mice. ( A – D ) Representative ( A ) H&E staining and ( C ) AB-PAS staining of colon tissues, and ( B , D ) the quantification of colonic injury area was performed using Image J software. In H&E staining, nuclei are stained blue–purple and cytoplasm appears pink; in AB-PAS staining, acidic mucins are stained blue and neutral mucins magenta. Scale bar = 100 μm. ( E , F ) Representative immunofluorescence co-staining images of Villin (green) and Ki67 (red), as well as Cd11b (green) and E-cadherin (red), in colon tissue. DAPI was used for nuclear labeling (blue). Scale bar = 100 μm. ( G ) Relative mRNA levels of barrier genes Muc2 , Zo1 , Ecad , and Ocln , stem/proliferation markers Lgr5 and Ki67 , and inflammatory genes Ccl2 , Il1b , Tnfα , and Il6 in colon tissues. Statistical significance: Data are presented as mean ± SD. The p values were calculated by one-way ANOVA followed by appropriate post hoc tests, as described in the . n = 6 mice per group. # p < 0.05, ## p < 0.01, ### p < 0.001 compared between the CT group and the model group. * p < 0.05, ** p < 0.01, *** p < 0.001 compared between the model group and the treatment groups.

Article Snippet: Sections were then blocked with 0.2% Triton X-100 + 2.5% BSA + 10% goat serum blocking buffer at RT for 30 min. After blocking, primary antibody cocktails were applied: Villin (Proteintech Group Inc., Rosemont, IL, USA; 66096-1-Ig) + Ki67 (Proteintech Group Inc., Rosemont, IL, USA; 27309-1-AP) or CD11b (Proteintech Group Inc., Rosemont, IL, USA; 66519-1-Ig) + E-cadherin (Cell Signaling Technology, Inc., Danvers, MA, USA; 3195S).

Techniques: Staining, Software, Immunofluorescence, Labeling

Journal: Cell & Bioscience

Article Title: Tie2-expressing monocytes/macrophages promote angiogenesis in chronically ischaemic brain tissue

doi: 10.1186/s13578-025-01401-1

Figure Lengend Snippet: Immunofluorescence and western blotting results showing the expression of relevant cytokines in the CIBT of 2VO + EMS rats. A Representative triple immunofluorescence staining images showing ANGPT2 + , Tie2 + , and CD11b + cells in the CIBT in each group (white arrows indicate TEMs). Bar = 50 µm. B Counts of ANGPT2 + cells in each group (n = 3, one-way ANOVA and Šídák's multiple comparisons test). C Counts of TEMs (Tie2 + and CD11b + ) in each group (n = 9, one-way ANOVA and Šídák's multiple comparisons test). D Representative western blot showing the relative expression of TRPS1, ANGPT2, Tie2, CD11b, VEGFA, IGF1, and CD31 in the CIBT adjacent to the TM tissue in each group (normalized to β-actin expression). E Densitometric analyses of the relative expression of TRPS1, ANGPT2, Tie2, CD11b, VEGFA, IGF1, and CD31 (n = 3, one-way ANOVA and Šídák's multiple comparisons test; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001). The error bars represent the ± SDs. CIBT: chronically ischaemic brain tissue; EMS: encephalomyosynangiosis; TEMs: Tie2-expressing monocytes/macrophages; 2VO: 2-vessel occlusion

Article Snippet: After being washed with PBST, the slices were incubated with 488-conjugated goat anti-rabbit (1:2000, ab150077, Abcam, UK), 594-conjugated goat anti-mouse (1:2000, ab150116, Abcam, UK), and 647-conjugated donkey anti-goat (1:2000, ab150135, Abcam, UK) secondary antibodies at room temperature in a dark room for 2 h. The 647-conjugated CD11b antibody (0.5 μg, CL647-65229, proteintech, China) was used to directly label CD11b.

Techniques: Immunofluorescence, Western Blot, Expressing, Staining

Journal: Cell & Bioscience

Article Title: Tie2-expressing monocytes/macrophages promote angiogenesis in chronically ischaemic brain tissue

doi: 10.1186/s13578-025-01401-1

Figure Lengend Snippet: Immunofluorescence results showing the expression of pro-angiogenesis factors in the CIBT of 2VO + EMS rats. A Representative triple immunofluorescence staining images showing Tie2 + , CD11b + , and VEGFA + cells and in the CIBT in each group (white arrows indicate TEMs). Bar = 50 µm. B Counts of VEGF + cells in each group (n = 3, one-way ANOVA and Šídák's multiple comparisons test). C Representative triple immunofluorescence staining images showing Tie2 + , CD11b + , and IGF1 + cells in the CIBT in each group (white arrows indicate TEMs). Bar = 50 µm. D Counts of IGF1 + cells in each group (n = 3, one-way ANOVA and Šídák's multiple comparisons test). CIBT: chronically ischaemic brain tissue; EMS: encephalomyosynangiosis; TEMs: Tie2-expressing monocytes/macrophages; 2VO: 2-vessel occlusion

Article Snippet: After being washed with PBST, the slices were incubated with 488-conjugated goat anti-rabbit (1:2000, ab150077, Abcam, UK), 594-conjugated goat anti-mouse (1:2000, ab150116, Abcam, UK), and 647-conjugated donkey anti-goat (1:2000, ab150135, Abcam, UK) secondary antibodies at room temperature in a dark room for 2 h. The 647-conjugated CD11b antibody (0.5 μg, CL647-65229, proteintech, China) was used to directly label CD11b.

Techniques: Immunofluorescence, Expressing, Staining