Journal: Translational Pediatrics

Article Title: Development and external validation of a mitophagy-related diagnostic model for biliary atresia based on cellular infiltration patterns

doi: 10.21037/tp-2025-1-864

Figure Lengend Snippet: Single cell sequencing and immunohistochemistry validate key molecular markers for BA. (A) Representative immunohistochemistry images validating the protein expression of mitophagy-related genes ( IGF2BP3 , NEDD4L , and ALDH2 ) in liver tissues from BA patients and controls (scale bar =100 µm). Ctrl images are from non-tumor liver tissue adjacent to hepatoblastoma specimens. Positive staining appears as brown discoloration and infiltrating immune cells were indicated by red arrows. (B) Differential analysis of single-cell transcription. (a) Single-cell’s cell classification landscape in all samples. (b) The single-cell expression differences of IGF2BP3, NEDD4L, PPIB, and ALDH2 between BA and Ctrl groups. BA, biliary atresia; Ctrl, control; FC, fold change.

Article Snippet: Next, sections were incubated overnight at 4 °C with the following primary antibodies: anti-human ALDH2 (HUABIO, Hangzhou, China; M1509-1, 1:800), NEDD4L (Proteintech, Wuhan, China; 13690-1-AP, 1:800), and IGF2BP3 (Proteintech, Wuhan, China; 14642-1-AP, 1:800).

Techniques: Single Cell, Sequencing, Immunohistochemistry, Expressing, Staining, Control

Journal: Translational Pediatrics

Article Title: Development and external validation of a mitophagy-related diagnostic model for biliary atresia based on cellular infiltration patterns

doi: 10.21037/tp-2025-1-864

Figure Lengend Snippet: The interaction between drugs and IGF2BP3 and NEDD4L proteins. (A-C) The interaction between cyclosporine and IGF2BP3 protein. (D-F) The interaction between cyclosporine and NEDD4L protein. Blue represents drug molecules, with red indicating oxygen atoms within the molecules. Green represents amino acid residues of the protein interacting with the drug molecules. The yellow background indicates the average interaction energy between the protein and drugs (average of 10 simulations).

Article Snippet: Next, sections were incubated overnight at 4 °C with the following primary antibodies: anti-human ALDH2 (HUABIO, Hangzhou, China; M1509-1, 1:800), NEDD4L (Proteintech, Wuhan, China; 13690-1-AP, 1:800), and IGF2BP3 (Proteintech, Wuhan, China; 14642-1-AP, 1:800).

Techniques:

Journal: Translational Pediatrics

Article Title: Development and external validation of a mitophagy-related diagnostic model for biliary atresia based on cellular infiltration patterns

doi: 10.21037/tp-2025-1-864

Figure Lengend Snippet: Potential schematic representation of the BA-CIMDGM genes in the pathogenesis of BA. Upregulated IGF2BP3 in T cells and NEDD4L in macrophages leads to impaired mitophagy and promotes immune activation and cytokines released. And the downregulation of ALDH2 in hepatocytes compromises the mitochondrial antioxidant defense and excessive ROS production, synergistically driving biliary epithelial cell injury and progressive liver fibrosis. BA, biliary atresia; BA-CIMDGM, Biliary Atresia Cell Infiltration Mitophagy Diagnostic Gene Model; ROS, reactive oxygen species.

Article Snippet: Next, sections were incubated overnight at 4 °C with the following primary antibodies: anti-human ALDH2 (HUABIO, Hangzhou, China; M1509-1, 1:800), NEDD4L (Proteintech, Wuhan, China; 13690-1-AP, 1:800), and IGF2BP3 (Proteintech, Wuhan, China; 14642-1-AP, 1:800).

Techniques: Activation Assay, Diagnostic Assay

Journal: Journal of pharmacological sciences

Article Title: Insulin-like growth factor 2 promotes osteoclastogenesis increasing inflammatory cytokine levels under hypoxia.

doi: 10.1016/j.jphs.2022.03.007

Figure Lengend Snippet: Fig. 1. Effect of IGF2 on osteoclastogenesis under normoxia and hypoxia. (A and B) Mouse BMCs were cultured under normoxia (20%) and supplemented with IGF2. Cells were fixed and stained for TRAP activity as a marker of osteoclastogenesis. Scale bar ¼ 1 mm. TRAP-positive (TRAP[þ]) multinucleated cells (MNCs) containing 3 nuclei were counted in each well. *P < 0.05 comparing control (0 ng/mL) and IGF2 treatment. Data represent the mean ± SEM (n ¼ 4 in each group). (C and D) Mouse BMCs were cultured under hypoxia (5%) and supplemented with IGF2. Cells were fixed and stained for TRAP activity as a marker of osteoclastogenesis. Scale bar ¼ 1 mm. TRAP[þ] MNCs containing 3 nuclei were counted in each well. *P < 0.05 comparing control (0 ng/mL) and IGF2 treatment. Data represent the mean ± SEM (n ¼ 4 in each group).

Article Snippet: Recombinant mouse IGF2 and mouse IGF2 antibodies were obtained from R&D Systems (Minneapolis, MN).

Techniques: Cell Culture, Staining, Activity Assay, Marker, Control

Journal: Journal of pharmacological sciences

Article Title: Insulin-like growth factor 2 promotes osteoclastogenesis increasing inflammatory cytokine levels under hypoxia.

doi: 10.1016/j.jphs.2022.03.007

Figure Lengend Snippet: Fig. 2. Effect of anti-IGF2 neutralizing antibody (a-IGF2 antibody) on osteoclastogenesis under normoxia and hypoxia. (A and B) Mouse BMCs were cultured under normoxia (20%) and supplemented with a-IGF2 antibody. Cells were fixed and stained for TRAP activity as a marker of osteoclastogenesis. Scale bar ¼ 1 mm. TRAP-positive (TRAP[þ]) multinu- cleated cells (MNCs) containing 3 nuclei were counted in each well. *P < 0.05 comparing control (0 ng/mL) and a-IGF2 antibody treatment. Data represent the mean ± SEM (n ¼ 4 in each group). (C and D) Mouse BMCs were cultured under hypoxia (5%) and supplemented with a-IGF2 antibody. Cells were fixed and stained for TRAP activity as a marker of osteoclastogenesis. Scale bar ¼ 1 mm. TRAP[þ] MNCs containing 3 nuclei were counted in each well. *P < 0.05 comparing control (0 ng/mL) and a-IGF2 antibody treatment. Data represent the mean ± SEM (n ¼ 4 in each group).

Article Snippet: Recombinant mouse IGF2 and mouse IGF2 antibodies were obtained from R&D Systems (Minneapolis, MN).

Techniques: Cell Culture, Staining, Activity Assay, Marker, Control

Journal: Journal of pharmacological sciences

Article Title: Insulin-like growth factor 2 promotes osteoclastogenesis increasing inflammatory cytokine levels under hypoxia.

doi: 10.1016/j.jphs.2022.03.007

Figure Lengend Snippet: Fig. 3. Effect of IGF2 stimulation on activation of the AkteNF-kB pathway. (A) Expression of Akt, phosphorylated Akt (Thr308 and Ser473), NF-kB, phosphorylated NF-kB (Ser536), and iNOS protein under 20% O2 conditions was analyzed by Western blotting. (B) Semi-quantitative analyses of protein expression are shown. Phosphorylated NF-kB and Akt levels were normalized to total NF-kB and total Akt levels, respectively. iNOS levels were normalized to actin levels. *P < 0.05 comparing control (0 ng/mL) and 100 ng/mL IGF2 treatment. Data represent the mean ± SEM (n ¼ 3 in each group).

Article Snippet: Recombinant mouse IGF2 and mouse IGF2 antibodies were obtained from R&D Systems (Minneapolis, MN).

Techniques: Activation Assay, Expressing, Western Blot, Control

Journal: Journal of pharmacological sciences

Article Title: Insulin-like growth factor 2 promotes osteoclastogenesis increasing inflammatory cytokine levels under hypoxia.

doi: 10.1016/j.jphs.2022.03.007

Figure Lengend Snippet: Fig. 4. Effect of IGF2 stimulation on expression levels of OC-related and inflammatory cytokine genes. (A) RT-qPCR analysis of the expression of selected genes in OCs treated with 100 ng/mL IGF2 under 20% O2 conditions. The relative mRNA expression level in the control (0 ng/mL) was defined as 1 or 100. *P < 0.05 comparing control (0 ng/mL) and 100 ng/ mL IGF2 treatment. Data represent the mean ± SEM (n ¼ 3 in each group). (B) RT-qPCR analysis of the expression of selected genes in non-OCs treated with 100 ng/mL IGF2 under 20% O2 conditions. The relative mRNA expression level in the control (0 ng/mL) was defined as 1 or 100. *P < 0.05 comparing control (0 ng/mL) and 100 ng/mL IGF2 treatment (n ¼ 3). Data represent the mean ± SEM (n ¼ 3 in each group).

Article Snippet: Recombinant mouse IGF2 and mouse IGF2 antibodies were obtained from R&D Systems (Minneapolis, MN).

Techniques: Expressing, Quantitative RT-PCR, Control

Journal: Oncotarget

Article Title: IGFBP2 modulates the chemoresistant phenotype in esophageal adenocarcinoma

doi:

Figure Lengend Snippet: A. Following 24-hour treatment with ON-TARGETplus IGFBP2 siRNA SMARTpool (si IGFBP2 pool), siNon-Targeting control (siNonT) or lipofectamine alone (Mock), Flo-1 cells were mock-treated or treated with 1 μg/mL (3.3 μM) CDDP, 2.5 μg/mL (8.3 μM) CDDP, 2.5 μg/mL (19.2 μM) 5-FU or 5 μg/mL (38.4 μM) 5-FU for 3 days and analyzed for cell viability using Cell Proliferation Reagent WST-1. Concurrent qRT-PCR was performed to verify IGFBP2 knockdown. B. Following 24-hour treatment with individual ON-TARGETplus IGFBP2 siRNAs, siNon-Targeting controls or lipofectamine alone, Flo-1 cells were mock-pretreated or pretreated with 200 ng/mL IGF1 or IGF2 for 1 hour followed by mock-treatment or treatment with 1 or 2.5 μg/mL (3.3 or 8.3 μM) CDDP in serum-free or 10% serum DMEM for 3 days. Cell viability was analyzed using Cell Proliferation Reagent WST-1. Columns and error bars are the mean ± SD of 3 or more wells in each experiment. Concurrent qRT-PCR was performed to verify IGFBP2 knockdown. (1 P, 1 μg/mL CDDP; 2.5 P, 2.5 μg/mL CDDP; 2.5 5-FU, 2.5 μg/mL 5-FU; 5 5-FU, 5 μg/mL 5-FU)

Article Snippet: Recombinant Human IGFBP2 (Cat#: 674-B2), IGF1 (Cat#: 291-G1), and IGF2 (Cat#: 292-G2) were purchased from R&D Systems (Minneapolis, MN).

Techniques: Control, Quantitative RT-PCR, Knockdown