d gal foxo4 dri group  (MedChemExpress)

Journal: Food Chemistry: Molecular Sciences

Article Title: Usage of nanobody-beta-galactosidase fusion in immunoassays and its application in detecting a peanut allergen

doi: 10.1016/j.fochms.2026.100357

Figure Lengend Snippet: Tagging a nanobody to the N-terminus of β-gal does not affect its activity. (A) Colonies of dH5α bacteria transformed with a plasmid containing the coding sequence for β-gal with Nb16 fused to its N-terminus via a 4AA (GSHV) linker and plated on an LB/Kan plate with IPTG and X-gal. The plate was photographed 24 h after incubation at 37 °C for 16 h. (B), (C), and (D) are the same as (A), except that the bacteria were induced to express β-gal with Nb16 fused to its N-terminus via a longer flexible peptide (GSGASGSHV), a Strep-tag-containing peptide (GSWSHPQFEKHV), or β-gal with purification tags, respectively.

Article Snippet: Milli-Q water was purified in-house using a Milli-Q Advantage A10 system (Millipore, Bedford, MA, USA) and used throughout. o -Nitrophenyl-β-galactoside (ONPG), Isopropyl β-D-1-thiogalactopyranoside (IPTG), Kanamycin (Kan), and X-Gal were purchased from GoldBio (St Louis, MO, USA).

Techniques: Activity Assay, Bacteria, Transformation Assay, Plasmid Preparation, Sequencing, Incubation, Strep-tag, Purification

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: FOXO4-DRI regulates endothelial cell senescence via the P53 signaling pathway

doi: 10.3389/fbioe.2025.1729166

Figure Lengend Snippet: FOXO4-DRI can effectively maintain vascular function in naturally aging mice. (A) Western blot experiments were conducted and quantitatively analyzed to determine the changes in protein levels of Ki-67, Lamin B, P21, P16, and γ-H2AX in the aortas of naturally aged mice after FOXO4-DRI or PBS treatment. n ≥ 4 per group. (B) RT-qPCR analysis was performed to examine the mRNA levels of Il-1β, Il-6, Cxcl15 , and Tnf-α in the aortas of naturally aged mice after FOXO4-DRI and PBS treatments. n = 3 per group. (C) Representative SA-β-Gal staining and quantitative analysis were performed to determine the number of positive cells in the aortas of naturally aged mice after FOXO4-DRI and PBS treatments. n = 5 per group. Scale bar = 100 μm. (D) Representative HE staining and quantitative analysis were performed to evaluate the thickness of the aortas in naturally aged mice after FOXO4-DRI and PBS treatments. n = 5 per group. Scale bar = 100 μm. (E) Color Doppler imaging and analysis were performed to detect and analyze the structural and blood flow conditions of the aortas in naturally aged mice after FOXO4-DRI and PBS treatments. n = 6 per group. (F) Representative DHE staining and quantitative analysis were performed to assess the ROS levels in the aortas of naturally aged mice after FOXO4-DRI and PBS treatments. n = 5 per group. Scale bar = 100 μm. Data are presented as mean ± SEM and analyzed using a two-tailed Student’s t-test. * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001.

Article Snippet: In the D-Gal + FOXO4-DRI group, mice also received D-galactose injections and, after 4 weeks, were given intraperitoneal injections of FOXO4-DRI (5 mg/kg, MCE, HY-P4157) every 2 days for 4 weeks, followed by tissue collection.

Techniques: Western Blot, Quantitative RT-PCR, Staining, Imaging, Two Tailed Test

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: FOXO4-DRI regulates endothelial cell senescence via the P53 signaling pathway

doi: 10.3389/fbioe.2025.1729166

Figure Lengend Snippet: FOXO4-DRI can effectively maintain vascular function in progeroid mice. (A) Western blot experiments were conducted and quantitatively analyzed to determine the changes in protein levels of Ki-67, Lamin B, P21, P16, and γ-H2AX in the aortas of progeroid mice after FOXO4-DRI or PBS treatment. n ≥ 4 per group. (B) RT-qPCR analysis was performed to examine the mRNA levels of Il-1β , Il-6 , Cxcl15 , and Tnf-α in the aortas of progeroid mice after FOXO4-DRI and PBS treatments. n = 3 per group. (C) Representative SA-β-Gal staining and quantitative analysis were performed to determine the number of positive cells in the aortas of progeroid mice after FOXO4-DRI and PBS treatments. Scale bar = 100 μm. n = 5 per group. (D) Representative HE staining and quantitative analysis were performed to evaluate the thickness in the aortas of progeroid mice after FOXO4-DRI and PBS treatments. Scale bar = 100 μm. n = 5 per group. (E) Color Doppler imaging and analysis were performed to detect and analyze the structural and blood flow conditions in the aortas of progeroid mice after FOXO4-DRI and PBS treatments. n = 5 per group. (F) Representative DHE staining and quantitative analysis were performed to assess the ROS levels in the aortas of progeroid mice after FOXO4-DRI and PBS treatments. n = 5 per group. Scale bar = 100 μm. Data are presented as mean ± SEM and analyzed using a two-tailed Student’s t-test. * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001.

Article Snippet: In the D-Gal + FOXO4-DRI group, mice also received D-galactose injections and, after 4 weeks, were given intraperitoneal injections of FOXO4-DRI (5 mg/kg, MCE, HY-P4157) every 2 days for 4 weeks, followed by tissue collection.

Techniques: Western Blot, Quantitative RT-PCR, Staining, Imaging, Two Tailed Test

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: FOXO4-DRI regulates endothelial cell senescence via the P53 signaling pathway

doi: 10.3389/fbioe.2025.1729166

Figure Lengend Snippet: FOXO4-DRI can alleviate endothelial cell senescence induced by OGD. (A) Western blot analysis was conducted to quantify the changes in protein levels of Ki-67, Lamin B, P21, P16, and γ-H2AX in senescent HUVECs following treatment with FOXO4-DRI or PBS, with n ≥ 4 per group. (B) Representative immunofluorescence analysis (green for Ki-67, blue for DAPI) was performed to assess the number of Ki-67 positive HUVECs in different groups, with n = 5 per group and scale bar = 40 μm. (C) Fluorescent staining was carried out to analyze the content and localization changes of P21 and γ-H2AX in different groups, with n = 5 per group and scale bar = 40 μm. (D) Representative SA-β-GAL staining and quantification were performed to evaluate the number of positive cells in different groups, with n = 5 per group and scale bar = 100 μm. (E) Tube formation assays were conducted to analyze relative tube lengths in different groups, with n = 5 per group and scale bar = 400 μm. (F) Representative DHE staining and quantification were carried out to evaluate the number of DHE-positive cells in different groups, with n = 5 per group and scale bar = 100 μm. (G) Scratch assays were performed to assess endothelial cell migration coverage at 0, 12, 24, and 36 h in different groups, with n = 5 per group and scale bar = 400 μm. Data are represented as mean ± SEM. Statistical analysis assessed by 2-way RM ANOVA. (H) RT-qPCR analysis was performed to detect mRNA levels of IL-6 , IL-8 , IL-1β , and TNF-α in different groups, with n = 3 per group. Data are presented as mean ± SEM and analyzed using a two-tailed Student’s t-test. * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001.

Article Snippet: In the D-Gal + FOXO4-DRI group, mice also received D-galactose injections and, after 4 weeks, were given intraperitoneal injections of FOXO4-DRI (5 mg/kg, MCE, HY-P4157) every 2 days for 4 weeks, followed by tissue collection.

Techniques: Western Blot, Immunofluorescence, Staining, Migration, Quantitative RT-PCR, Two Tailed Test

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: FOXO4-DRI regulates endothelial cell senescence via the P53 signaling pathway

doi: 10.3389/fbioe.2025.1729166

Figure Lengend Snippet: FOXO4-DRI can block the interaction between P53 and FOXO4. (A) The FOXO4-P53 interaction was analyzed through co-immunoprecipitation in OGD-treated HUVECs, in different groups, with n = 3 per group. (B) Western blot analysis quantified changes in protein levels of pSer46-P53 and P53 in different groups, with n = 5 per group. (C) Western blot analysis quantified changes in protein levels of pSer46-P53 and P53 in different groups, with n = 5 per group. (D) Western blot analysis was performed to examine the distribution of pSer46-P53 and P53 in the cytoplasm and nucleus of HUVECs in different groups. With n = 5 per group. (E) Subcellular localization of pSer46-P53 in different groups of HUVECs was analyzed using representative immunofluorescence staining (pSer46-P53 in green, DAPI in blue), with n = 5 per group, scale bar = 40 μm. (F) Immunofluorescence detection and analysis of subcellular localization of pSer46-P53 and FOXO4 in aortic sections of naturally aged mice (pSer46-P53 in green, FOXO4 in red, DAPI in blue), with n = 5 per group, scale bar = 20 μm. (G) Immunofluorescence detection and analysis of subcellular localization of pSer46-P53 and FOXO4 in aortic sections of progeroid mice (pSer46-P53 in green, FOXO4 in red, DAPI in blue), with n = 5 per group, scale bar = 20 μm. Data are presented as mean ± SEM and analyzed using a two-tailed Student’s t-test. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: In the D-Gal + FOXO4-DRI group, mice also received D-galactose injections and, after 4 weeks, were given intraperitoneal injections of FOXO4-DRI (5 mg/kg, MCE, HY-P4157) every 2 days for 4 weeks, followed by tissue collection.

Techniques: Blocking Assay, Immunoprecipitation, Western Blot, Immunofluorescence, Staining, Two Tailed Test

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: FOXO4-DRI regulates endothelial cell senescence via the P53 signaling pathway

doi: 10.3389/fbioe.2025.1729166

Figure Lengend Snippet: FOXO4-DRI promotes apoptosis in senescent cells. (A) Western blot analysis quantified changes in protein levels of BAX, Bcl2, and c-Caspase3 in different groups, with n = 5 per group. (B) Western blot analysis assessed changes in protein levels of BAX, Bcl2, and c-Caspase3 in the aortas of naturally aged mice after FOXO4-DRI or PBS treatment, with n = 5 per group. (C) Representative TUNEL staining and quantitative analysis were conducted to determine the number of positive cells in the aortas of naturally aged mice after FOXO4-DRI and PBS treatments, with n = 5 per group and scale bar = 100 μm. (D) Representative TUNEL staining and quantification evaluated the number of TUNEL-positive cells across different groups with n = 5 per group and scale bar = 20 μm. (E) Flow cytometry analysis was used to quantitatively assess the number of Annexin V-positive cells in different groups, with a sample size of n = 5 per group. Data are presented as mean ± SEM and analyzed using a two-tailed Student’s t-test. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: In the D-Gal + FOXO4-DRI group, mice also received D-galactose injections and, after 4 weeks, were given intraperitoneal injections of FOXO4-DRI (5 mg/kg, MCE, HY-P4157) every 2 days for 4 weeks, followed by tissue collection.

Techniques: Western Blot, TUNEL Assay, Staining, Flow Cytometry, Two Tailed Test

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: FOXO4-DRI regulates endothelial cell senescence via the P53 signaling pathway

doi: 10.3389/fbioe.2025.1729166

Figure Lengend Snippet: Schematic illustration of FOXO4-DRI in alleviating endothelial cell senescence aging extends cell survival. This study demonstrates that FOXO4-DRI mitigates senescence by inhibiting the FOXO4-P53 interaction, promoting P53 phosphorylation, and inducing apoptosis in senescent cells, thereby alleviating aging-related symptoms. Content produced with BioRender; license VN28TABHO4.

Article Snippet: In the D-Gal + FOXO4-DRI group, mice also received D-galactose injections and, after 4 weeks, were given intraperitoneal injections of FOXO4-DRI (5 mg/kg, MCE, HY-P4157) every 2 days for 4 weeks, followed by tissue collection.

Techniques: Phospho-proteomics, Produced

Journal: mBio

Article Title: Loss of LafB activity reverses daptomycin resistance in E. faecium

doi: 10.1128/mbio.00715-25

Figure Lengend Snippet: TLC analysis of glycolipids in total membrane lipid extracts from E. faecium . Glycolipid profiles of E. faecium SM1, SM1 lafB T577C, SM1Δ lafB , and SM1 Δ lafB /pAT28:: lafB mutants were compared to WT E. faecium SM1. Purified lipid standards included Gal₂DAG [1,2-diacyl-3-O-(α-D-galactosyl(1→6)-β-D-galactosyl)-sn-glycerol] and Glc ₁ DAG [1,2-diacyl-3-O-(α-D-glucopyranosyl)-sn-glycerol] (Avanti Polar Lipids). Glycolipid species Glc ₁ DAG, Glc ₂ DAG, and Gal ₂ DAG were identified based on migration relative to standards. Additional bands representing unknown lipid species are also indicated. The absence of glycolipid bands in the Δ lafB mutant and their restoration in the complemented strain demonstrates the role of lafB in glycolipid biosynthesis. Data are representative of three independent experiments.

Article Snippet: Approximately 5 μL of each of the two standards (Gal 2 DAG: 1,2-diacyl-3- O -(α- d -galactosyl1-6)-β- d -galactosyl- sn -glycerol Avanti Polar Lipids 840524P, Glc 1 DAG: 1,2-diacyl-3-O-(α-D-glucopyranosyl)-sn-glycerol Avanti Polar Lipids 840522P) and ~15 μL of each sample was spotted on the baseline of a Silica gel 60 TLC plate.

Techniques: Membrane, Purification, Migration, Mutagenesis