Journal: Journal of experimental & clinical cancer research : CR

Article Title: Regnase-1 downregulation promotes pancreatic cancer through myeloid-derived suppressor cell-mediated evasion of anticancer immunity.

doi: 10.1186/s13046-023-02831-w

Figure Lengend Snippet: Fig. 6 Suppression of CTLs by CD11b+ MDSCs is responsible for the acceleration of tumor progression by Regnase-1 downregulation. A-D Evaluation of phenotypes of orthotopic syngeneic tumors of WT or Regnase-1 KO murine pancreatic cancer cells. Representative macro images of pancreatic tumors (A). Relative mRNA levels of Cd8a, Ifng, Fasl, and Gzmb (B) (N = 6 per group). Representative images of HE (C, left panel) and CD8a immunostaining (C, right panel) and the number of CD8-positive cells (C, right) (N = 6 per group). Dot plots of CD3+CD8+ cells evaluated by flow cytometry (D, left) and the proportion of CD8 + cells among CD45+ cells (D, right) (N = 3 per group). E–H Evaluation of phenotypes of orthotopic syngeneic tumors of WT or Regnase-1-KO murine pancreatic cancer cells with or without depletion of CD8+ cells upon anti-CD8a antibody or IgG treatment. Experimental schematic (E). Dot plots of CD3+ and CD8.+ cells in WT or Regnase-1-KO syngeneic tumors upon anti-CD8a antibody or IgG treatment evaluated by flow cytometry (F). Tumor weights (G) (N = 6 per group). The relative mRNA levels of Cd8a, Ifng, Fasl, and Gzmb (H) (N = 6 per group). Student’s t test was used to evaluate differences between 2 groups. One-way ANOVA with Tukey’s post hoc test was used to compare differences among 4 groups. *P < 0.05, scale bars: 100 μm (insets)

Article Snippet: BE0061, a fully neutralizing mAb recognizing CD8a, and control IgG were obtained from Bioxcell.

Techniques: Immunostaining, Flow Cytometry

Journal: Oncology Reports

Article Title: SNX10 regulates the proliferation, apoptosis and cell cycle of acute B lymphoblastic leukemia cells via the PI3K/Akt signaling pathway

doi: 10.3892/or.2025.8911

Figure Lengend Snippet: SNX10 promotes B-ALL cellular behaviors via the PI3K/Akt signaling pathway. (A) Venn diagram showing the 211 significantly differentially expressed proteins identified between SNX10-OE Rs4;11 and EV cells. (B) Volcano plot of the differentially expressed proteins showed 76 downregulated and 135 upregulated proteins. (C) KEGG pathway enrichment analysis of the differentially expressed proteins. SNX10, sorting nexin 10; B-ALL, B-cell acute lymphoblastic leukemia; OE, overexpression; EV, empty vector; KEGG, Kyoto Encyclopedia of Genes and Genomes.

Article Snippet: Idelalisib (IDEL, TargetMoi, Cas: 870281-82-6) is a small molecule inhibitor that targets the PI3K catalytic subunit p110δ.

Techniques: Analysis, Over Expression, Plasmid Preparation, Significance Assay

Journal: Oncology Reports

Article Title: SNX10 regulates the proliferation, apoptosis and cell cycle of acute B lymphoblastic leukemia cells via the PI3K/Akt signaling pathway

doi: 10.3892/or.2025.8911

Figure Lengend Snippet: SNX10 influences the activity of the PI3K/Akt signaling pathway in B-ALL cells. The protein expression levels of PI3K, p-PI3K, Akt and p-Akt proteins in SNX10-OE and knockdown (A) Rs4;11 and (B) Nalm-6 cells were detected. After treating SNX10-OE B-ALL cells with 10 µM IDEL (a PI3K inhibitor) for 24 h, the protein expression levels of PI3K, p-PI3K, Akt and p-Akt proteins in SNX10-OE and knockdown (C) Rs4;11 and (D) Nalm-6 cells were detected. Data are presented as the mean ± SD of three independent repeats and compared using a one-way or two-way ANOVA. *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001 vs. EV group or shNC group. SNX10, sorting nexin 10; B-ALL, B-cell acute lymphoblastic leukemia; OE, overexpression; EV, empty vector; shRNA, short hairpin RNA; NC, negative control; IDEL, Idelalisib; p-, phosphorylated.

Article Snippet: Idelalisib (IDEL, TargetMoi, Cas: 870281-82-6) is a small molecule inhibitor that targets the PI3K catalytic subunit p110δ.

Techniques: Activity Assay, Expressing, Knockdown, Over Expression, Plasmid Preparation, shRNA, Negative Control, Western Blot

Journal: Oncology Reports

Article Title: SNX10 regulates the proliferation, apoptosis and cell cycle of acute B lymphoblastic leukemia cells via the PI3K/Akt signaling pathway

doi: 10.3892/or.2025.8911

Figure Lengend Snippet: SNX10 affects the activity of B-ALL cells through the PI3K/Akt signaling pathway. After treating (A) Rs4;11 cells and (B) Nalm-6 cells with 10 µM IDEL (a PI3K inhibitor) for 24 h, cell proliferation was analyzed using a Cell Counting Kit-8 assay. Apoptosis of (C) Rs4;11 cells and (D) Nalm-6 cells (annexin V/7-AAD staining) was analyzed by flow cytometry. Cell cycle distribution in (E) Rs4;11 cells and (F) Nalm-6 cells was analyzed by flow cytometry. Data are presented as the mean ± SD of three independent repeats and compared using a one-way or two-way ANOVA. ***P<0.001 and ****P<0.0001 vs. EV group. SNX10, sorting nexin 10; B-ALL, B-cell acute lymphoblastic leukemia; OE, overexpression; EV, empty vector; IDEL, Idelalisib; ns, not significant.

Article Snippet: Idelalisib (IDEL, TargetMoi, Cas: 870281-82-6) is a small molecule inhibitor that targets the PI3K catalytic subunit p110δ.

Techniques: Activity Assay, Cell Counting, Staining, Flow Cytometry, Over Expression, Plasmid Preparation, FACS

Journal: Journal of Ovarian Research

Article Title: Gut microbiota-derived Tryptophanol driven by N-Carbamylglutamate alleviates premature ovarian failure through inhibiting oxidative stress

doi: 10.1186/s13048-026-02152-4

Figure Lengend Snippet: Tryptophanol treatment alleviated POF symptoms by inhibiting PI3K/AKT and MAPK pathways. A Experimental scheme. B Estrus status of mice ( n = 8). C Body weights of mice ( n = 8). D Ovary weights of mice ( n = 8). E Representative Ovary from the Control, the POF and the NCG groups mice with H&E and IHC staining. H&E scale bar = 200 μm; IHC scale bar = 100 μm. F Quantification of primordial, primary, secondary, antral follicles, corpora lutea and corpora atretica per ovary. n = 3 ovaries per each condition. Levels of FSH ( G ), LH ( H ), E 2 ( I ) in mouse serum ( n = 6). Levels of ROS ( J ), MDA ( K ), SOD ( L ), GSH-Px ( M ) in mouse ovaries ( n = 3–4). N Western blotting ( a ) and quantitative ( b ) analysis of proteins ( n = 3). O Litter sizes of mice ( n = 3). Data presented as the mean ± SD, statistical significance was determined using one-way or two-way ANOVA. * p < 0.05, ** p < 0.01. *** p < 0.001. Different lowercase letters indicate significant differences ( p < 0.05)

Article Snippet: The membrane was blocked with 5% bovine serum albumin (BSA; Sigma) for 1 h at room temperature, followed by overnight incubation at 4 °C with the primary antibodies: rabbit Cleaved caspase 3 (MCE, HY- P80623 , 1:800), rabbit BAX (Bioworld, BS61098, 1:800), rabbit BCL2 (Bioworld, BS1511, 1:800), rabbit ERK1/2 (Bioworld, BS90472, 1:1,000), rabbit p-ERK1/2 (Bioworld, BS4621P, 1:1,000), rabbit p38 (Bioworld, BS4635, 1:1,000), rabbit p-p38 (Bioworld, BS4766, 1:1,000), rabbit PI3K (MCE, HY- P80867 , 1:1,000); rabbit p-PI3K (MCE, HY- P80846 , 1:1,000), rabbit AKT (Bioworld, MB3288 1:1,000); rabbit p-AKT (Bioworld, BS4007 1:1,000); rabbit IL-6 (Abclonal, A0286, 1:800), rabbit IL-1β (MCE, HY- P80720 , 1:1,000), rabbit Occludin (Abclonal, A2601, 1:1,000); rabbit ZO-1 (Bioworld, BS71522, 1:1,000); rabbit GPX4 (MCE, HY- P80450 , 1:1,000); mouse GAPDH (Bioworld, MB001, 1:1,000); rabbit SOD2 (Bioworld, BS6734, 1:1,000).

Techniques: Control, Immunohistochemistry, Western Blot

Journal: Journal of Ovarian Research

Article Title: Gut microbiota-derived Tryptophanol driven by N-Carbamylglutamate alleviates premature ovarian failure through inhibiting oxidative stress

doi: 10.1186/s13048-026-02152-4

Figure Lengend Snippet: Tryptophanol treatment alleviated POF by inhibiting PI3K/AKT and MAPK pathways in vitro . A Experimental scheme. B Different concentrations of Cy on GCs viability was assessed by CCK8 assay. C and E Different concentrations of Trp on GCs viability was assessed by CCK8 assay. D The effect of Trp on GCs cell viability following Cy treatment. The control and treatment groups were stained with DCFH-DA, the cells were washed and examined by fluorescence microscopy. Scale bar = 50 μm. F The fluorescence intensity of ROS in mouse GCs. G The level of MDA in mouse GCs. H Flow cytometry apoptosis analysis graph and cell death rate in different groups. The level of SOD ( I ), GSH-Px ( J ), and E 2 ( K ) in mouse GCs. ( L ) Western blotting ( a ) and quantitative ( b ) analysis of proteins ( n = 3). All values are presented as the mean ± SD ( n = 3). Different lowercase letters indicate significant differences compared to the control group ( p < 0.05)

Article Snippet: The membrane was blocked with 5% bovine serum albumin (BSA; Sigma) for 1 h at room temperature, followed by overnight incubation at 4 °C with the primary antibodies: rabbit Cleaved caspase 3 (MCE, HY- P80623 , 1:800), rabbit BAX (Bioworld, BS61098, 1:800), rabbit BCL2 (Bioworld, BS1511, 1:800), rabbit ERK1/2 (Bioworld, BS90472, 1:1,000), rabbit p-ERK1/2 (Bioworld, BS4621P, 1:1,000), rabbit p38 (Bioworld, BS4635, 1:1,000), rabbit p-p38 (Bioworld, BS4766, 1:1,000), rabbit PI3K (MCE, HY- P80867 , 1:1,000); rabbit p-PI3K (MCE, HY- P80846 , 1:1,000), rabbit AKT (Bioworld, MB3288 1:1,000); rabbit p-AKT (Bioworld, BS4007 1:1,000); rabbit IL-6 (Abclonal, A0286, 1:800), rabbit IL-1β (MCE, HY- P80720 , 1:1,000), rabbit Occludin (Abclonal, A2601, 1:1,000); rabbit ZO-1 (Bioworld, BS71522, 1:1,000); rabbit GPX4 (MCE, HY- P80450 , 1:1,000); mouse GAPDH (Bioworld, MB001, 1:1,000); rabbit SOD2 (Bioworld, BS6734, 1:1,000).

Techniques: In Vitro, CCK-8 Assay, Control, Staining, Fluorescence, Microscopy, Flow Cytometry, Western Blot

Journal: Theranostics

Article Title: CCR2 deficiency protects against doxorubicin-induced cardiac dysfunction through enhanced IL12B-dependent autophagy

doi: 10.7150/thno.131005

Figure Lengend Snippet: IL12B is upregulated in CCR2-deficient hearts and activates autophagy through PI3K/Akt/mTOR pathway inhibition. (A) Western blot results of IL12A and IL12B protein in heart from WT and CCR2 -/- mouse (n = 6). (B) ELISA quantification of IL12A and IL12B levels in heart lysates from WT and CCR2 -/- mouse (n = 6). (C) Western blot results of IL12B protein in BMDMs differentiated from WT and CCR2 -/- mouse (n = 3 independent experiments). (D) Immunofluorescence co-staining of IL12B (green) and the macrophage marker CD68 (red) in heart sections from WT and CCR2 -/- mouse. Scale bar: 5 μm. (E) Western blot results of autophagy markers (LC3B-II and P62) in H9C2 cells treated with recombinant IL12B protein (n = 3 independent experiments). (F) Quantitative PCR analysis of autophagy-related genes Atg12, Atg5, Atg7, Atg3, and Gabarapl1 in H9C2 cells following vehicle or IL12B treatment (2 ng/mL) (n = 3 independent experiments). (G) Western blot results of autophagy-related signaling pathway in H9C2 treated with vehicle or IL12B (2 ng/mL) (n = 3 independent experiments). (H) Western blot results of pPI3K, PI3K and autophagy markers (LC3B-II, ATG12, GABARAPL1, and P62) in H9C2 treated with vehicle, IL12B (2 ng/mL) alone, or in combination with the PI3K activator 740 Y-P (3 μM) (n = 3 independent experiments).

Article Snippet: To measure antibody titers in mouse serum by indirect ELISA, 96-well plates were coated with 1 μg/mL recombinant IL12A or IL12B protein (MCE, HY-P700099AF and HY-P700100AF).

Techniques: Inhibition, Western Blot, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Marker, Recombinant, Real-time Polymerase Chain Reaction

Journal: Theranostics

Article Title: CCR2 deficiency protects against doxorubicin-induced cardiac dysfunction through enhanced IL12B-dependent autophagy

doi: 10.7150/thno.131005

Figure Lengend Snippet: IL12B promotes cardiomyocyte autophagy in vitro . (A) Cell viability assay of H9C2 treated with DOX (1 μM) in the presence or absence of recombinant IL12B proteins (2 ng/mL) (n = 3 independent experiments). (B) Western blot analysis of autophagy markers (LC3B-II, ATG12, GABARAPL1, and P62) in the same groups (n = 3 independent experiments). (C) Immunofluorescence staining and quantitative analysis of LC3B puncta in the same groups (n = 3 independent experiments). Scale bar: 20 μm. (D) Tandem fluorescence microscopy of H9C2 cells expressing the mRFP-GFP-LC3 reporter. Yellow puncta represent autophagosomes, and red puncta represent autolysosomes. Scale bar: 10 μm (n = 3 independent experiments). (E) Transmission electron microscopy of H9C2. Arrows indicate autolysosomes. Scale bar: 2 μm, 0.5 μm (n = 3 independent experiments).

Article Snippet: To measure antibody titers in mouse serum by indirect ELISA, 96-well plates were coated with 1 μg/mL recombinant IL12A or IL12B protein (MCE, HY-P700099AF and HY-P700100AF).

Techniques: In Vitro, Viability Assay, Recombinant, Western Blot, Immunofluorescence, Staining, Fluorescence, Microscopy, Expressing, Transmission Assay, Electron Microscopy

Journal: Theranostics

Article Title: CCR2 deficiency protects against doxorubicin-induced cardiac dysfunction through enhanced IL12B-dependent autophagy

doi: 10.7150/thno.131005

Figure Lengend Snippet: Recombinant IL12B administration protects against DIC through autophagy activation in vivo . (A) Experimental design for in vivo IL12B administration. (B) Representative M-mode echocardiographic tracings of mouse from the vehicle, DOX alone, and DOX + IL12B groups. Scale bar: 0.1 s, 2 mm. (C, D) Quantitative analysis of EF and FS in the indicated groups (n = 6). (E–G) The levels of cardiac injury markers in serum, including LDH-L, CK-MB, and cTnI (n = 6). (H) Western blot analysis of autophagy markers (LC3B-II, ATG12, GABARAPL1, and P62) in heart tissues from the same groups (n = 6). (I) Quantitative PCR analysis of autophagy-related gene (Atg12, Atg5, Atg7, Atg3, and Gabarapl1) in heart tissues from the same groups (n = 6). (J) Immunofluorescence staining and quantitative analysis of LC3B puncta in cardiomyocytes from the same groups. Scale bar: 50 μm (n = 6). (K) Transmission electron microscopy analysis of autolysosome density from the same groups. Arrows indicate autolysosomes. Scale bars: 1 μm and 0.5 μm (n = 6).

Article Snippet: To measure antibody titers in mouse serum by indirect ELISA, 96-well plates were coated with 1 μg/mL recombinant IL12A or IL12B protein (MCE, HY-P700099AF and HY-P700100AF).

Techniques: Recombinant, Activation Assay, In Vivo, Western Blot, Real-time Polymerase Chain Reaction, Immunofluorescence, Staining, Transmission Assay, Electron Microscopy

Journal: Theranostics

Article Title: CCR2 deficiency protects against doxorubicin-induced cardiac dysfunction through enhanced IL12B-dependent autophagy

doi: 10.7150/thno.131005

Figure Lengend Snippet: IL12B neutralization abolishes CCR2 deficiency-mediated cardioprotection in vivo . (A) Experimental design of IL12B neutralization in CCR2 -/- mouse. (B) Representative M-mode echocardiographic tracings of CCR2 -/- mouse treated with vehicle, DOX alone, or DOX + anti-IL12B Neu. Scale bar: 0.1 s, 2 mm. (C, D) Quantitative results of EF and FS in the indicated groups (n = 6). (E–G) The levels of cardiac injury markers in serum, including LDH-L, CK-MB, and cTnI (n = 6). (H) Western blot results of autophagy markers (LC3B-II, ATG12, GABARAPL1, and P62) in heart tissues from CCR2 -/- mouse treated with vehicle, DOX alone, or DOX + anti-IL12B Neu (n = 6). (I) Quantitative PCR analysis of autophagy-related gene (Atg12, Atg5, Atg7, Atg3, and Gabarapl1) in heart tissues from the same groups (n = 6). (J) Immunofluorescence staining and quantitative analysis of LC3B puncta in cardiomyocytes from the same groups. Scale bar: 50 μm (n = 6). (K) Transmission electron microscopy analysis of autolysosome density in cardiomyocytes from the same groups. Arrows indicate autolysosomes. Scale bars: 1 μm and 0.5 μm (n = 6).

Article Snippet: To measure antibody titers in mouse serum by indirect ELISA, 96-well plates were coated with 1 μg/mL recombinant IL12A or IL12B protein (MCE, HY-P700099AF and HY-P700100AF).

Techniques: Neutralization, In Vivo, Western Blot, Real-time Polymerase Chain Reaction, Immunofluorescence, Staining, Transmission Assay, Electron Microscopy

Journal: Theranostics

Article Title: CCR2 deficiency protects against doxorubicin-induced cardiac dysfunction through enhanced IL12B-dependent autophagy

doi: 10.7150/thno.131005

Figure Lengend Snippet: Pharmacological CCR2 inhibition protects against DIC without compromising the antitumor efficacy of doxorubicin in 4T1 breast cancer model. (A) Cell viability assay of 4T1 breast cancer cells treated with DOX or in combination with CCR2 antagonist4 (n = 3 independent experiments). (B, C) Tumor growth curves and representative tumor images at the study endpoint in a syngeneic 4T1 breast cancer model. Scale bar: 1 cm (n = 6). (D) Representative M-mode echocardiographic tracings of tumor bearing mouse treated with vehicle, DOX alone, CCR2 antagonist4 alone or DOX + CCR2 antagonist4. Scale bar: 0.1 s, 2 mm. (E, F) Quantitative results of EF and FS in the same groups (n = 6). (G–I) The levels of cardiac injury markers in serum, including LDH-L, CK-MB, and cTnI (n = 6). (J) Western blot results of autophagy markers (LC3B-II, ATG12, GABARAPL1, and P62) in heart tissues from the same groups (n = 6). (K) Quantitative PCR analysis of autophagy-related gene (Atg12, Atg5, Atg7, Atg3, and Gabarapl1) in heart tissues from the same groups (n = 6). (L) Working model. DOX administration triggers cardiomyocyte dysfunction, ultimately leading to heart failure. In contrast, loss of function of CCR2 initiates a protective cascade. Specifically, CCR2 deficiency increases the secretion of IL12B. This cytokine directly targets cardiomyocytes, suppressing PI3K/Akt/mTOR signaling to trigger protective autophagy, which clears damaged organelles and toxic proteins, thus sustaining cellular homeostasis and cardiac function.

Article Snippet: To measure antibody titers in mouse serum by indirect ELISA, 96-well plates were coated with 1 μg/mL recombinant IL12A or IL12B protein (MCE, HY-P700099AF and HY-P700100AF).

Techniques: Inhibition, Viability Assay, Western Blot, Real-time Polymerase Chain Reaction