antibody against cd44  (Bioss)

Journal: Frontiers in Veterinary Science

Article Title: PDGFD maintains ovine tail ADSCs in a proliferative state by suppressing CXCL8 and activating PI3K/MAPK signaling

doi: 10.3389/fvets.2026.1777426

Figure Lengend Snippet: Immunofluorescence identification of CD44 in ovine ADSCs. (a) CD44 immunofluorescence staining (red) shows strong positive expression localized to the cell membrane and cytoplasm. (b) DAPI staining (blue) marks the cell nuclei. (c) Merged image illustrates the subcellular localization of CD44.

Article Snippet: To reduce non-specific binding, cells were blocked with 1% bovine serum albumin (BSA, Bioss, Beijing, China) at room temperature for 30 min. After blocking, cells were incubated overnight at 4 °C with a primary antibody against CD44 (rabbit polyclonal antibody, Bioss, Cat No. bs-55039R, dilution 1:1,000).

Techniques: Immunofluorescence, Staining, Expressing, Membrane

Journal: Journal of Nanobiotechnology

Article Title: Hyaluronic acid-based reduction responsive nanoparticles for Improved anti-psoriasis effects of traditional Chinese herb monomer oleanolic acid via blocking YAP-AREG axis

doi: 10.1186/s12951-026-04264-x

Figure Lengend Snippet: The in vitro anti-psoriasis effects of OA-NPs@OA. ( A ) CLSM images and ( B ) Flow cytometry data of the cellular uptake of free Cy5.5 (for OA-substitution, 0.5 µg/mL) and Cy5.5-loaded OA-NPs (OA-NPs@Cy5.5, 0.5 µg/mL Cy5.5) in HaCaT cells incubated for different times. Bar in ( A ) = 50 μm. ( C ) IF analysis of CD44 expression in HaCaT cells with or without LPS-stimulation by CLSM. Bar = 100 μm. ( D ) Relative fluorescence intensity of CD44. ( E ) Flow cytometric analysis of OA-NPs@Cy5.5 uptake (Cy5.5: 0.5 µg/mL) by HaCaT cells stimulated with LPS for different durations. ( F ) Anti-proliferation effects of HaCaT cells at various concentrations (based on OA: 0.01, 0.1, 0.5, 1, 2, 5, 10, 20, 50 µg/mL) of free OA, OA-NPs, and OA-NPs@OA and their IC 50 values, as determined by MTT assays. ( G ) Statistical significance analysis of IC 50 values of free OA, OA-NPs, and OA-NPs@OA. ( H ) Analysis of apoptosis percentages in HaCaT cells treated with OA and OA-NPs@OA for 24 h and 48 h by flow cytometry with Annexin V/Propidium Iodide (PI) staining. ( I ) Representative crystal violet staining of HaCaT cells treated with free OA or OA-NPs@OA for 24 h and 48 h. Bar = 500 μm. **** p < 0.0001, n.s. = no significance

Article Snippet: The sections were incubated overnight at 4 °C with the primary antibody against CD44 (Servicebio, GB112054 , 1:300 dilution), followed by a 50-min incubation at room temperature with the corresponding secondary antibody (Servicebio, GB25303, 1:400 dilution).

Techniques: In Vitro, Flow Cytometry, Incubation, Expressing, Fluorescence, Staining

Journal: Journal of Nanobiotechnology

Article Title: Hyaluronic acid-based reduction responsive nanoparticles for Improved anti-psoriasis effects of traditional Chinese herb monomer oleanolic acid via blocking YAP-AREG axis

doi: 10.1186/s12951-026-04264-x

Figure Lengend Snippet: Drug Deposition Study of OA-NPs@OA in mice psoriatic lesions after topical administration. ( A ) CLSM images showing the fluorescence distribution of topically applied OA-NPs@Cy5.5 and free Cy5.5 (1 µg per mouse) in psoriatic lesions after 2–24 h.‌ Bar = 100 μm. ( B ) Quantitative analysis of fluorescence intensity in psoriatic lesions (from Fig. 3A). ( C ) Fluorescence distribution of topically applied OA-NPs@Cy5.5 and free Cy5.5 in the epidermis and dermis of psoriatic lesions at 2–24 h post-administration (from Fig. 3A). ( D ) CD44 IF staining and OA-NPs@Cy5.5 fluorescence images in psoriatic skin lesions and normal skin of mice showing their co-localization and the ( E ) quantification values. Bar = 100 μm. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, n.s. = no significance

Article Snippet: The sections were incubated overnight at 4 °C with the primary antibody against CD44 (Servicebio, GB112054 , 1:300 dilution), followed by a 50-min incubation at room temperature with the corresponding secondary antibody (Servicebio, GB25303, 1:400 dilution).

Techniques: Fluorescence, Staining

Journal: bioRxiv

Article Title: ASPARAGINE-RICH METASTATIC NICHES DRIVE PROSTATE CANCER ORGANOTROPISM BY ENABLING TRANSLATIONAL REWIRING TOWARD N-GLYCOSYLATED PROTEINS

doi: 10.64898/2026.02.27.708521

Figure Lengend Snippet: (A) Western blot analysis of ASNS in metastatic cells isolated from bone (B-M-1, B-M-2) and lung (L-M-1, L-M-2) lesions. Vinculin was used as loading control. The image is representative of three independent experiments. (B) Schematic representation of asparaginyl-tRNA synthetase 1 (NARS1) mechanism of action. (C) NARS1 mRNA levels in PC3 cells following NARS1 silencing. Cells were transfected with NARS1-targeting small interfering RNA (siRNA) or negative control, and mRNA levels were evaluated after 5 days of incubation in 3D cultures (3D-C) by quantitative RT-PCR, using scramble-transfected cells as reference. (D) Relative cell number of PC3 cells silenced for NARS1 and cultured under standard 2D conditions for 5 days in the presence or absence of Asn (0.1 mM). One-way ANOVA with Sidak’s correction. (E-G) Number of putative N-glycosylation sites in proteins encoded by genes up- or down-regulated in metastatic cells derived from bone (E), lung (F), and liver (G) relative to primary tumor (PT). RNA-seq analysis was conducted as described in . Values are expressed relative to total protein number. (H-I) Fractional enrichment of UDP-GlcNAc isotopologues. PC3 cells were grown in 2D or 3D-C for 5 days and subsequently incubated in a medium containing U- 13 C-glucose for 24h. Labeling enrichment was evaluated by LC-MS analysis, and isotopologue abundance is reported as relative to total UDP-GlcNAc amount. Welch’s t test. (J) Labeling (m+5) enrichment of penotose phosphates from U- 13 C-glucose in PC3 cells growing in 2D or 3D-C for 5 days and subsequently incubated in a medium containing U- 13 C-glucose for 24h. Labeling enrichment was evaluated by LC-MS analysis. (K, L, P) Western blot analysis of GFPT1 (K), STT3a (L), and CD44 (P) expression in PC3 cells silenced for GFPT1, STT3a/b, and CD44 respectively after 48h of gene silencing. Vinculin was used as a loading control. The image is representative of three independent experiments. (M) Concanavalin A lectin binding assay performed on lysates from PC3 cells silenced or not for GFPT1 or STT3a/b and cultured in 3D-C for 5 days. Immunoblot for vinculin was used to confirm equal protein loading across samples. The image is representative of three independent experiments. (N-O) Adhesion of PC3 3D-C to collagen type I (L) and hyaluronic acid (M). Cells were cultured with Asn (0,1 mM) for 5 days and allowed to adhere for 15 min to plates coated with matrix components as reported. Adherent cells were quantified and data are shown relative to untreated cells. Welch’s t-test.

Article Snippet: Cells were permeabilized with 0.25% Triton X-100 in PBS for 5 minutes, washed twice with PBS, and incubated overnight at 4°C with primary antibody against CD44 (Cell Signaling Technology #3570) diluted 1:100 in IFF.

Techniques: Western Blot, Isolation, Control, Transfection, Small Interfering RNA, Negative Control, Incubation, Quantitative RT-PCR, Cell Culture, Glycoproteomics, Derivative Assay, RNA Sequencing, Labeling, Liquid Chromatography with Mass Spectroscopy, Expressing, Binding Assay

Journal: bioRxiv

Article Title: ASPARAGINE-RICH METASTATIC NICHES DRIVE PROSTATE CANCER ORGANOTROPISM BY ENABLING TRANSLATIONAL REWIRING TOWARD N-GLYCOSYLATED PROTEINS

doi: 10.64898/2026.02.27.708521

Figure Lengend Snippet: (A) Volcano plot showing differential protein abundance in PC3 cells grown in 3D-C in the absence or presence of Asn (0.1 mM). The x-axis reports the difference (log 2 fold change) fold change between 3D-C and 3D-C+Asn; the y-axis shows –log 10 (p value). Proteins specific to 3D-C were identified by excluding proteins also expressed in 2D conditions. Each dot represents a quantified protein; red dots denote proteins exceeding the ≥2-fold change threshold. (B) Western blot analysis of CD44 expression in metastatic cells isolated from bone (B-M-1, B-M-2) and lung (L-M-1, L-M-2) lesions isolated as described in . Vinculin was used as a loading control. Representative of three independent experiments. (C) Western blot analysis of CD44 in PC3 cells cultured under standard 2D conditions or in 3D-C. Vinculin was used as a loading control. Representative of three independent experiments. (D) Time-course Western blot analysis of CD44 expression in PC3 cells cultured in 3D-C during spheroid formation (5 h, 1 day, 2 days, and 5 days after plating). Vinculin was used as a loading control. The image is representative of three independent experiments. (E) Representative immunohistochemical staining for CD44 in primary prostate carcinoma tissues (PT) and bone metastatic (BM) lesions from patients with PC. Boxed areas indicate regions shown at higher magnification. Brown staining denotes CD44-positive cells; nuclei are stained with hematoxylin. (F) H-score quantification of CD44 immunohistochemical staining shown in (E). H-scores were calculated by integrating staining intensity and the percentage of positive cells. Welch’s t test. (G) Total spheroid area of CD44-silenced PC3 cells grown in 3D-C compared to control silencing condition. Welch’s t-test. (H) Hyaluronic acid adhesion assay on CD44-silenced PC3 cells grown in 3D-C for 5days before plating on hyaluronic acid-coated plate. Data are shown relative to untreated cells. Welch’s t test. (I) Western blot analysis of CD44 expression in PC3 cells cultured in 3D-C in the presence or absence of Asn (0.1 mM and 1 mM). Vinculin was used as a loading control. The image is representative of three independent experiments. (J) Western blot analysis of CD44 in PC3 cells in 3D-C with or without Asn (0.1 mM) in the presence or absence of L-asparaginase (ASNase, 0.25 U/ml). Vinculin was used as loading control. The image is representative of three independent experiments. (K) CD44 mRNA expression levels in PC3 cells cultured in 3D-C with or without Asn (0.1 mM), measured by quantitative RT-PCR and normalized to the non-treated condition. Student’s t test. (L) Confocal fluorescence microscopy images showing CD44 membrane localization in PC3 cells grown in standard 2D conditions, 3D-C, and 3D-C supplemented with Asn (0,1 mM). The image is representative of three independent experiments. (M, N) Western blot analysis of CD44 expression in PC3 cells grown in 3D-C and silenced for NARS1 (M) or GFPT1 (N), cultured with or without Asn (0.1 mM). Vinculin was used as a loading control. The images are representative of three independent experiments. (O) Total spheroid area of 3D-C PC3 cells silenced for the CD44 with or without Asn (0.1 mM). One-way ANOVA with Dunnett’s correction. (P) Hyaluronic acid adhesion assay on CD44-silenced PC3 cells grown in 3D-C with or without Asn (0.1 mM). One-way ANOVA with Dunnett’s correction. ns, not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Data represent mean ± s.e.m. from at least three independent experiments.

Article Snippet: Cells were permeabilized with 0.25% Triton X-100 in PBS for 5 minutes, washed twice with PBS, and incubated overnight at 4°C with primary antibody against CD44 (Cell Signaling Technology #3570) diluted 1:100 in IFF.

Techniques: Quantitative Proteomics, Western Blot, Expressing, Isolation, Control, Cell Culture, Immunohistochemical staining, Staining, Cell Adhesion Assay, Quantitative RT-PCR, Fluorescence, Microscopy, Membrane