anti human foxo1 rabbit mab  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: OBESITY-INDUCED ENDOTHELIAL FENESTRATION AND CAPILLARY LEAKAGE CONTRIBUTE TO INCREASED PAIN SENSATION

doi: 10.64898/2026.03.13.711502

Figure Lengend Snippet: (A) Schematic diagrams illustrate the mechanisms involved in the NGF production in response to insulin signaling activation in keratinocytes (left) and the sensory hypersensitivity mediated by the NGF-TrkA-PI3K signaling pathway in sensory nerves (right). Insulin stimulates NGF expression in keratinocytes , accompanied by FOXO1 translocation from the nucleus to the cytoplasm. The NGF-TrkA-PI3K signaling pathway is involved in sensory hypersensitivity, likely by sensitizing the TRPV1 channel and facilitating its trafficking to the plasma membrane . “C” indicates the cytoplasm; “N” indicates the nucleus. (B) Representative images of immunofluorescence staining of cultured human keratinocytes, labeled with an antibody to NGF (green or white), along with the nuclear marker TOPRO3 (red), are presented. Note that non-specific nuclear signals were observed when using the anti-NGF antibody for immunostaining. Scale bars: 20 μm. (C) Quantification of cytoplasmic signals using the anti-NGF antibody staining in human keratinocytes cultured with or without insulin is shown. N=3 in each group. (D) Representative images of immunofluorescence staining of cultured human keratinocytes, labeled with an antibody to FOXO1 (green or white), along with the nuclear marker TOPRO3 (red), are presented. The dotted box regions in the middle panels are magnified in the right panels. Scale bars: 20 μm. (E) Quantification of nuclear signals using the anti-FOXO1 antibody staining in human keratinocytes cultured with or without insulin is shown. N=3 in each group. (F) Quantification of cytoplasmic signals using the anti-FOXO1 antibody staining in human keratinocytes cultured with or without insulin is shown. N=3 in each group. (G) Representative images of immunofluorescence staining of human keratinocytes cultured with or without insulin or si FOXO1 , labeled with the anti-NGF antibody (green or white), along with TOPRO3 (red), are presented. (H) Quantification of cytoplasmic signals using the anti-NGF antibody staining in human keratinocytes cultured with or without insulin or si FOXO1 is shown. N=3 in each group. Results are shown as the mean ± SEM. **p<0.01, ***p<0.001. P values were determined by the parametric two-tailed t test. The schematic diagrams and graphic summary were partially created with BioRender.com .

Article Snippet: Staining was performed using rabbit anti-FOXO1 antibody (Cell Signaling, 2880) and guinea pig anti-K14 antibody (PROGEN, GP-CK14, 1:100).

Techniques: Activation Assay, Expressing, Translocation Assay, Clinical Proteomics, Membrane, Immunofluorescence, Staining, Cell Culture, Labeling, Marker, Immunostaining, Two Tailed Test

Journal: bioRxiv

Article Title: OBESITY-INDUCED ENDOTHELIAL FENESTRATION AND CAPILLARY LEAKAGE CONTRIBUTE TO INCREASED PAIN SENSATION

doi: 10.64898/2026.03.13.711502

Figure Lengend Snippet: (A) Representative section immunohistochemical images of ear skin from control mice, DIO mice treated with saline, and DIO mice treated with the neutralizing anti-PLVAP antibody are presented. This assay uses the antibodies for FOXO1 (green), the keratinocyte marker K14 (red), along with the nuclear marker TOPRO3 (blue). Each inset displays the pattern of FOXO1 expression in a single keratinocyte. Dashed lines indicate the boundary between the epidermis and the dermis. “Epi” indicates the epidermis; “D” indicates the dermis. Scale bars: 20 μm. (B) Quantification of nuclear FOXO1 expression in keratinocytes is provided. The percentages of nuclear FOXO1 expression within the total FOXO1 expression in keratinocytes are presented. The sample sizes are as follows: N = 5 in control, N = 5 in DIO + Saline, N = 7 in DIO + PLVAP Ab. (C) Representative X-gal staining images of ear skin from NGF-LacZ control mice, DIO mice with saline, and DIO mice with the neutralizing anti-PLVAP antibody (blue) are presented. Dashed lines indicate the boundary between the epidermis and the dermis. Scale bars: 50 μm. (D) Quantification of the LacZ-positive area in the epidermis is provided. The sample sizes are as follows: N = 9 in control, N = 15 in DIO + Saline, N = 9 in DIO + PLVAP Ab. (E) Graphical summary illustrates how vascular hyperpermeability leads to sensory hypersensitivity in DIO skin. Increased permeability in the superficial dermal capillaries facilitates the diffusion of insulin into the epidermis, activating insulin signaling in epidermal keratinocytes. This activation leads to NGF upregulation in these keratinocytes, which in turn promotes sensory hypersensitivity in DIO skin. A neutralizing anti-PLVAP antibody reduces the diffusion of insulin, thereby decreasing NGF expression in the epidermal keratinocytes and alleviating sensory hypersensitivity. Results are shown as the mean ± SEM. *p<0.05, **p<0.01. P values were determined by the parametric two-tailed t test. The schematic diagrams and graphic summary were partially created with BioRender.com .

Article Snippet: Staining was performed using rabbit anti-FOXO1 antibody (Cell Signaling, 2880) and guinea pig anti-K14 antibody (PROGEN, GP-CK14, 1:100).

Techniques: Immunohistochemical staining, Control, Saline, Marker, Expressing, Staining, Permeability, Diffusion-based Assay, Activation Assay, Two Tailed Test