primary rabbit anti hcftr  (R&D Systems)

Journal: Clinical and Translational Radiation Oncology

Article Title: MonoHER selectively enhances the radiotherapy response in p53 wild-type breast cancer via stabilization of p53

doi: 10.1016/j.ctro.2026.101147

Figure Lengend Snippet: Effect of monoHER (M) combined with radiation (RT) on γ-H2AX immunofluorescence staining (at 24 h post-irradiation time point) in breast cancer and normal cells. Data are presented as mean ± SEM from three independent experiments (20 cells/experiment). *p < 0.05, **p < 0.01.

Article Snippet: Cells were incubated with a rabbit anti-human γH2AX primary antibody (1:500, Merck, JBW301), followed by detection using goat anti-rabbit Alexa Fluor 488 (1:500, Invitrogen, 11001).

Techniques: Immunofluorescence, Staining, Irradiation

Journal: Veterinary Sciences

Article Title: Clinical Spatial Distribution of Aquaporin-1 in Camel Cornea Using Assistive AI Applications

doi: 10.3390/vetsci13050425

Figure Lengend Snippet: Representative photomicrographs of the camel cornea. Panels (C1, MD1, MV1) show H&E-stained sections illustrating epithelial thickness in the central (C), middle dorsal (MD), and middle ventral (MV) regions (scale bar: 100 µm). Panels (C2, MD2, MV2) depict the stromal layer and Descemet’s membrane in the same regions following H&E staining. Panels (C3, MD3, MV3) demonstrate AQP1 immunoreactivity within the corneal epithelium and keratocytes of the anterior stroma, with variable staining intensity across regions (black arrows; scale bar: 50 µm). Panels (C4, MD4, MV4) show AQP1 localization in the posterior stroma and endothelium, where immunostaining is primarily confined to keratocytes and endothelial cells (black arrows; scale bar: 50 µm).

Article Snippet: Subsequently, the sections were incubated for 60 min with a rabbit polyclonal anti-human AQP1 primary antibody (1:1000; catalog no. GB11310, Servicebio, Woburn city, MA, USA).

Techniques: Staining, Membrane, Immunostaining

Journal: Veterinary Sciences

Article Title: Clinical Spatial Distribution of Aquaporin-1 in Camel Cornea Using Assistive AI Applications

doi: 10.3390/vetsci13050425

Figure Lengend Snippet: Representative photomicrographs of the camel cornea from the middle nasal (MN), middle temporal (MT), and peripheral dorsal (PD) regions. Panels (MN1, MT1, PD1) show H&E-stained sections illustrating epithelial thickness in the corresponding regions. In addition, vascular structures are visible in the peripheral dorsal region (PD2), likely associated with the limbal area (black arrows; scale bar: 100 µm). Panels (MN2, MT2, PD2) demonstrate the stromal layer and Descemet’s membrane in these regions following H&E staining. Panels (MN3, MT3, PD3) reveal AQP1 immunoreactivity within the corneal epithelium and keratocytes of the anterior stroma, with regional variation in staining intensity (black arrows; scale bar: 50 µm). Panels (MN4, MT4, PD4) illustrate AQP1 localization in the posterior stroma and endothelium, where staining is predominantly confined to keratocytes and endothelial cells (black arrows; scale bar: 50 µm).

Article Snippet: Subsequently, the sections were incubated for 60 min with a rabbit polyclonal anti-human AQP1 primary antibody (1:1000; catalog no. GB11310, Servicebio, Woburn city, MA, USA).

Techniques: Staining, Membrane

Journal: Veterinary Sciences

Article Title: Clinical Spatial Distribution of Aquaporin-1 in Camel Cornea Using Assistive AI Applications

doi: 10.3390/vetsci13050425

Figure Lengend Snippet: Representative photomicrographs of the camel cornea from the peripheral ventral (PV), peripheral nasal (PN), and peripheral temporal (PT) regions. Panels (PV1, PN1, PT1) show H&E-stained sections illustrating epithelial thickness in the respective regions (scale bar: 100 µm). Vascular structures are evident in the peripheral areas (PV2, PN2, PT2), likely corresponding to extensions of the limbal vasculature (black arrows). Panels (PV2, PN2, PT2) further demonstrate the stromal layer and Descemet’s membrane following H&E staining. Panels (PV3, PN3, PT3) display AQP1 immunoreactivity within the corneal epithelium and keratocytes of the anterior stroma, with noticeable regional differences in staining intensity (black arrows; scale bar: 50 µm). The strongest epithelial expression of AQP1 was observed in the peripheral nasal region (PN3), highlighted by white circles. Panels (PV4, PN4, PT4) illustrate AQP1 localization in the posterior stroma and endothelium, where staining is primarily confined to keratocytes and endothelial cells (black arrows; scale bar: 50 µm). Additionally, panel (PT5) shows the presence of brown melanin granules within the peripheral temporal region (black arrows; scale bar: 50 µm).

Article Snippet: Subsequently, the sections were incubated for 60 min with a rabbit polyclonal anti-human AQP1 primary antibody (1:1000; catalog no. GB11310, Servicebio, Woburn city, MA, USA).

Techniques: Staining, Membrane, Expressing

Journal: Veterinary Sciences

Article Title: Clinical Spatial Distribution of Aquaporin-1 in Camel Cornea Using Assistive AI Applications

doi: 10.3390/vetsci13050425

Figure Lengend Snippet: Immunohistochemical localization of AQP1 in camel corneal epithelium across different cellular layers, including superficial, intermediate (polyhedral), and basal cells. The columns represent the relative expression levels of AQP1 in the following corneal regions according to Area Fraction (%): central (C), middle dorsal (MD), middle nasal (MN), middle temporal (MT), middle ventral (MV), peripheral dorsal (PD), peripheral nasal (PN), peripheral temporal (PT), and peripheral ventral (PV). Data are presented as Mean ± SD (n = 6). Different superscript letters above bars indicate statistically significant differences between groups (One-way ANOVA followed by Tukey’s post hoc test, p < 0.05).

Article Snippet: Subsequently, the sections were incubated for 60 min with a rabbit polyclonal anti-human AQP1 primary antibody (1:1000; catalog no. GB11310, Servicebio, Woburn city, MA, USA).

Techniques: Immunohistochemical staining, Expressing

Journal: Veterinary Sciences

Article Title: Clinical Spatial Distribution of Aquaporin-1 in Camel Cornea Using Assistive AI Applications

doi: 10.3390/vetsci13050425

Figure Lengend Snippet: Immunohistochemical distribution of AQP1 in the camel cornea, including the anterior and posterior stromal regions as well as the endothelium. The columns illustrate the relative expression levels of AQP1 across different corneal regions according to Area Fraction (%): central (C), middle dorsal (MD), middle nasal (MN), middle temporal (MT), middle ventral (MV), peripheral dorsal (PD), peripheral nasal (PN), peripheral temporal (PT), and peripheral ventral (PV). Data are presented as Mean ± SD (n = 6). Different superscript letters above bars indicate statistically significant differences between groups (One-way ANOVA followed by Tukey’s post hoc test, p < 0.05).

Article Snippet: Subsequently, the sections were incubated for 60 min with a rabbit polyclonal anti-human AQP1 primary antibody (1:1000; catalog no. GB11310, Servicebio, Woburn city, MA, USA).

Techniques: Immunohistochemical staining, Expressing

Journal: Veterinary Sciences

Article Title: Clinical Spatial Distribution of Aquaporin-1 in Camel Cornea Using Assistive AI Applications

doi: 10.3390/vetsci13050425

Figure Lengend Snippet: Proposed model for the spatial distribution of AQP1 water channels in the camel cornea in the three corneal layers, epithelium, stroma and endothelium. The green color shows AQP1 localization in the different corneal epithelial cell layers; superficial, polyhedral, and basal cell layers. The black color shows localization of AQP1 in keratocyte cells of stroma, while the red color clarifies the localization of AQP1 in corneal endothelium.

Article Snippet: Subsequently, the sections were incubated for 60 min with a rabbit polyclonal anti-human AQP1 primary antibody (1:1000; catalog no. GB11310, Servicebio, Woburn city, MA, USA).

Techniques:

Journal: Veterinary Sciences

Article Title: Clinical Spatial Distribution of Aquaporin-1 in Camel Cornea Using Assistive AI Applications

doi: 10.3390/vetsci13050425

Figure Lengend Snippet: Topographical map of AQP1 distribution across the nine corneal regions. The schematic represents the regional intensity of AQP1 expression in the epithelium (EPI), stroma (STR), and endothelium (EN) of the camel cornea. The AI-generated Area Fraction (AF %) data: (+) = Weak expression (AF < 2%), (++) = Moderate expression (AF = 2–4%), (+++) = Strong expression (AF = 4–6%) and (++++) = Very strong expression (AF > 6%).

Article Snippet: Subsequently, the sections were incubated for 60 min with a rabbit polyclonal anti-human AQP1 primary antibody (1:1000; catalog no. GB11310, Servicebio, Woburn city, MA, USA).

Techniques: Expressing, Generated

Journal: STAR Protocols

Article Title: Protocol for generation of human gallbladder epithelia cells and their derived hepatocytes using a chemically defined approach

doi: 10.1016/j.xpro.2025.104302

Figure Lengend Snippet: Production and identification of human gallbladder epithelial cells (hGBECs) (A) Schematic diagram for the procedure of production and identification of hGBECs. (B) q-RT-PCR for expression of KRT19, EPCAM, LGR5, SOX9, HNF4A and ALB in each passage of hGBECs. Fold chang is calculated by 2 -ΔΔCt , relative to GAPDH (n= 3). Data are represented as mean ± SD. (C) Immunostaining for HNF4A, SOX9 (red), TROP2 and EPCAM (green) of hGBECs. Cell nuclei were stained by DAPI (blue). Scale bar = 100 μm.

Article Snippet: Anti-human SOX9 rabbit primary antibody (1:200 IF) , Merck , Cat#HPA001758; RRID: AB_1080067.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Immunostaining, Staining

Journal: STAR Protocols

Article Title: Protocol for generation of human gallbladder epithelia cells and their derived hepatocytes using a chemically defined approach

doi: 10.1016/j.xpro.2025.104302

Figure Lengend Snippet: Quality tests of hGBECs (Passage 3) (A) Growth curves of hGBECs at different densities (n=3). Data are represented as mean ± SD. (B) Table displaying sterility test results of hGBECs. (C) Flow cytometric analysis showing the proportion of SOX9+, TROP2+, HNF4A+, EPCAM+ cells. (D) Representative karyotype of Passage 3 cells.

Article Snippet: Anti-human SOX9 rabbit primary antibody (1:200 IF) , Merck , Cat#HPA001758; RRID: AB_1080067.

Techniques: Sterility

Journal: Clinical and Translational Radiation Oncology

Article Title: Differential regulation of radioadaptation by quercetin between human normal and cancer cells

doi: 10.1016/j.ctro.2025.101099

Figure Lengend Snippet: Quercetin restores NRF2 nuclear translocation in radioadapted MCF10A cells. Representative immunofluorescence images of NRF2 (green) and DAPI (blue) and quantification of the nuclear-to-cytoplasmic NRF2 fluorescence ratio in MCF10A cells 24 h after 5 Gy irradiation with or without prior LDRT and quercetin treatment (Scale bar: 10 μm). Data is shown as mean ± SEM. * p < 0.05, ** p < 0.01.

Article Snippet: Cells were incubated with rabbit anti-human NRF2 primary antibody (1:200; Proteintech, 16396–1-AP), and detection was performed using goat anti-rabbit Alexa Fluor 488 (1:500, Invitrogen, 11001).

Techniques: Translocation Assay, Immunofluorescence, Fluorescence, Irradiation

Journal: Research

Article Title: Targeting a Shared Mitophagy Regulator: The SIRT1–FOXO3–DEPP1 Axis Underpins the Dual Bone and Brain Benefits of Total Flavonoids from Drynaria fortunei

doi: 10.34133/research.1125

Figure Lengend Snippet: TFDF reverses a cross-tissue stress–autophagy signature and highlights DEPP1 as a shared node. (A) Venn diagram of TFDF-reversal DEGs in bone and the hippocampus (reversal defined as Model vs. Sham significant change, directionally opposed in TFDF-H vs. Model). (B) Pathway enrichment of reversal genes highlighting FOXO signaling (database and statistics in Materials and Methods). (C) Cross-filtering for Model↑ and TFDF-H↓ genes in both tissues identifies DEPP1 as a shared, treatment-reversed transcript (expression changes shown for bone and hippocampus). (D) Complementary enrichment of the reversal sets prioritizes FOXO signaling in both tissues. (E) GSEA plots demonstrating negative enrichment of FOXO programs in the Model vs. Sham groups and a positive shift in TFDF-H vs. Model (bone and hippocampus) groups.

Article Snippet: IF was performed on mouse hippocampal sections and on MC3T3-E1 and HT22 cells using primary antibodies DEPP1 (CUSABIO, CSB-PA865135LA01HU), LC3 (Immunoway, PT0235R), and TOM20 (Immunoway, PT0287R).

Techniques: Expressing

Journal: Research

Article Title: Targeting a Shared Mitophagy Regulator: The SIRT1–FOXO3–DEPP1 Axis Underpins the Dual Bone and Brain Benefits of Total Flavonoids from Drynaria fortunei

doi: 10.34133/research.1125

Figure Lengend Snippet: TFDF reduces DEPP1 expression and normalizes the expression of autophagy–mitochondrial markers in the hippocampus and bone of OVX–CUMS mice. (A) Representative hippocampal immunofluorescence images of NeuN (neurons, red) and DEPP1 (green) in the Sham, Model, TFDF-L, and TFDF-H groups; nuclei are stained with DAPI (blue). (B) Quantification of the DEPP1/NeuN double-positive area in the hippocampus. (C) Representative DEPP1 immunohistochemistry in trabecular bone. (D) Quantification of the DEPP1-positive area in bone. (E and F) Representative Western blots for SIRT1, FOXO3, and DEPP1 in the hippocampus (E) and bone (F). (G and H) Densitometric analysis of SIRT1, FOXO3, and DEPP1 expression in the hippocampus (G) and bone (H) normalized to that of β-actin. (I) Representative hippocampal immunofluorescence for NeuN (red) and LC3B (green). (J) Quantification of the hippocampal LC3B/NeuN double-positive area. (K) Representative LC3 immunohistochemistry in trabecular bone. (L) Quantification of the LC3-positive area in bone. (M and N) Western blots for p62, LC3B, and TOM20 in the hippocampus (M) and bone (N). (O and P) Densitometric analysis of p62, LC3B, and TOM20 expression in the hippocampus (O) and bone (P), normalized to that of β-actin.

Article Snippet: IF was performed on mouse hippocampal sections and on MC3T3-E1 and HT22 cells using primary antibodies DEPP1 (CUSABIO, CSB-PA865135LA01HU), LC3 (Immunoway, PT0235R), and TOM20 (Immunoway, PT0287R).

Techniques: Expressing, Immunofluorescence, Staining, Immunohistochemistry, Western Blot

Journal: Research

Article Title: Targeting a Shared Mitophagy Regulator: The SIRT1–FOXO3–DEPP1 Axis Underpins the Dual Bone and Brain Benefits of Total Flavonoids from Drynaria fortunei

doi: 10.34133/research.1125

Figure Lengend Snippet: TFDF normalizes the SIRT1–FOXO3–DEPP1 axis, rebalances excessive autophagy, and rescues mitochondrial and osteogenic functions in MC3T3-E1 cells. MC3T3-E1 cells were assigned to Control, Model (H 2 O 2 injury), TFDF, or NAC (positive antioxidant control) groups. Immunoblotting revealed that SIRT1 down-regulation, FOXO3 hyperacetylation, and DEPP1 up-regulation after H 2 O 2 were reversed by TFDF (A), as determined by densitometry (B). TMRE microscopy and quantification revealed ΔΨm loss in the Model group and restoration by TFDF (C and D). Autophagy/mitochondrial markers indicated over-autophagy under injury that were recalibrated toward baseline by TFDF (E and F). TEM revealed autophagosome accumulation and swollen mitochondria in the Model group, which were mitigated by TFDF (G). Multicolor IF (LC3B/TOM20/DEPP1) revealed increased LC3B puncta, TOM20 fragmentation, and DEPP1 elevation in the Model group, all of which improved with TFDF (H); colocalization analyses confirmed normalization of LC3B–TOM20 (mitophagy coupling) and a reduction in TOM20–DEPP1 coupling by TFDF (I and J). Osteogenic function assays demonstrated TFDF-mediated recovery of ALP activity and ARS mineral deposition (K), accompanied by increased RUNX2 expression and OCN/ALP expression, as determined by Western blotting (L and M). Data are presented as the mean ± SEM, with n indicated on the plots; the statistical tests and multiple-comparison procedures are described in Materials and Methods. Abbreviations: TFDF, total flavonoids of Drynaria fortunei ; NAC, N-acetyl-L-cysteine; TMRE, tetramethylrhodamine ethyl ester; LC3, microtubule-associated protein 1 light chain 3; OCN, osteocalcin.

Article Snippet: IF was performed on mouse hippocampal sections and on MC3T3-E1 and HT22 cells using primary antibodies DEPP1 (CUSABIO, CSB-PA865135LA01HU), LC3 (Immunoway, PT0235R), and TOM20 (Immunoway, PT0287R).

Techniques: Control, Western Blot, Microscopy, Activity Assay, Expressing, Comparison

Journal: Research

Article Title: Targeting a Shared Mitophagy Regulator: The SIRT1–FOXO3–DEPP1 Axis Underpins the Dual Bone and Brain Benefits of Total Flavonoids from Drynaria fortunei

doi: 10.34133/research.1125

Figure Lengend Snippet: TFDF mitigates oxidative injury in HT22 cells by restoring SIRT1–FOXO3–DEPP1 signaling and autophagy–mitochondrial homeostasis, thereby improving neuroplasticity. Cells were assigned to Control, Model (H 2 O 2 ), TFDF, or NAC (antioxidant control) groups. Western blots showing SIRT1–FOXO3–DEPP1↑ after injury and reversal by TFDF (A and B). TMRE imaging revealed ΔΨm loss in the Model group and rescue by TFDF (C and D). Autophagy/mitochondrial markers were recalibrated toward baseline by TFDF (E and F), which is consistent with TEM showing fewer autophagosomes and preserved cristae (G). IF (DAPI/LC3B/TOM20/DEPP1) demonstrated reduced LC3B puncta, increased TOM20 integrity, and decreased DEPP1 with TFDF (H), which was supported by colocalization readouts (DEPP1–TOM20 and LC3B–TOM20) (I and J). Quantification of DEPP1–TOM20 colocalization, and the mitophagy index is shown in (K). TFDF further increased the expression of BDNF and p-CREB/CREB and restored the expression of synapsin I and PSD-95 (L and M). Data are presented as the mean ± SEM; statistics and replicate numbers are provided in Materials and Methods. Abbreviations: TFDF, total flavonoids of Drynaria fortunei ; NAC, N-acetyl-L-cysteine; TMRE, tetramethylrhodamine ethyl ester.

Article Snippet: IF was performed on mouse hippocampal sections and on MC3T3-E1 and HT22 cells using primary antibodies DEPP1 (CUSABIO, CSB-PA865135LA01HU), LC3 (Immunoway, PT0235R), and TOM20 (Immunoway, PT0287R).

Techniques: Control, Western Blot, Imaging, Expressing

Journal: Research

Article Title: Targeting a Shared Mitophagy Regulator: The SIRT1–FOXO3–DEPP1 Axis Underpins the Dual Bone and Brain Benefits of Total Flavonoids from Drynaria fortunei

doi: 10.34133/research.1125

Figure Lengend Snippet: DEPP1 bidirectionally alters autophagy–mitochondrial coupling under oxidative injury in HT22 and MC3T3-E1 cells. DEPP1 expression was reduced by siRNA (KD) or increased by plasmid (OE); cells were exposed to H 2 O 2 to model injury. Protein and mRNA assays confirmed effective KD/OE in both lines (A and B). TMRE imaging revealed ΔΨm loss in the Model group, partial recovery in the KD+Model group, and a further decrease in the OE+Model group (C and D). TEM revealed swollen mitochondria and autophagosomes in the Model group, fewer autophagosomes after KD, and abundant autophagosomes after OE (E). Western blots demonstrated model-associated LC3-II accumulation, p62 depletion, and TOM20 reduction; KD shifted these toward control, whereas OE intensified them (F) with densitometry in (G). LC3B/TOM20 immunofluorescence revealed parallel changes in puncta burden and mitochondrial network integrity, as shown by the statistical data in (J) (H to J). Group labels: Control, Model, KD+Model, and OE+Model. Data are presented as the mean ± SEM; replicate numbers and statistics are provided in Materials and Methods.

Article Snippet: IF was performed on mouse hippocampal sections and on MC3T3-E1 and HT22 cells using primary antibodies DEPP1 (CUSABIO, CSB-PA865135LA01HU), LC3 (Immunoway, PT0235R), and TOM20 (Immunoway, PT0287R).

Techniques: Expressing, Plasmid Preparation, Imaging, Western Blot, Control, Immunofluorescence

Journal: Research

Article Title: Targeting a Shared Mitophagy Regulator: The SIRT1–FOXO3–DEPP1 Axis Underpins the Dual Bone and Brain Benefits of Total Flavonoids from Drynaria fortunei

doi: 10.34133/research.1125

Figure Lengend Snippet: DEPP1 knockdown with TFDF maintains and often augments pathway activity and functional rescue in injured osteoblasts and neurons. (A) Schematic of the gene–drug design and hypothesized placement of DEPP1 downstream of TFDF-responsive signaling. (B) Autophagy/mitochondrial Western blots (LC3-I/II, p62, and TOM20) in MC3T3-E1 and HT22 cells under Control, Model, siDEPP1+Model, TFDF+Model, and siDEPP1+TFDF+Model conditions. (C) Densitometry for panel (B). (D) Representative ROS fluorescence micrographs. (E) Quantification of intracellular ROS levels. (F) Osteogenic function of MC3T3-E1 cells: ALP staining (day 7) and ARS mineralization (days 14 to 21). (G) Osteogenic markers (RUNX2, OCN, and ALP) were measured by Western blotting. (H) Corresponding quantification. (I) Neuronal plasticity markers in HT22 cells (BDNF, p-CREB/CREB, Synapsin I, and PSD-95) were measured by Western blotting. (J) Quantification. In both cell types, siDEPP1 and TFDF each improved the injury phenotype, and siDEPP1+TFDF achieved comparable or greater improvement without occluding the effects of TFDF. Statistical tests and n values are provided in Materials and Methods and on the plots.

Article Snippet: IF was performed on mouse hippocampal sections and on MC3T3-E1 and HT22 cells using primary antibodies DEPP1 (CUSABIO, CSB-PA865135LA01HU), LC3 (Immunoway, PT0235R), and TOM20 (Immunoway, PT0287R).

Techniques: Knockdown, Activity Assay, Functional Assay, Western Blot, Control, Fluorescence, Staining

Journal: Research

Article Title: Targeting a Shared Mitophagy Regulator: The SIRT1–FOXO3–DEPP1 Axis Underpins the Dual Bone and Brain Benefits of Total Flavonoids from Drynaria fortunei

doi: 10.34133/research.1125

Figure Lengend Snippet: Docking and functional perturbation support SIRT1 as a key TFDF-responsive effector. (A) Docked pose of naringenin in the SIRT1 activator pocket with key hydrogen-bond and hydrophobic contacts indicated. (B) Radius of gyration, (C) number of protein–ligand hydrogen bonds, (D) backbone RMSF, (E) solvent-accessible surface area, and (F) ligand RMSD, all indicating rapid stabilization after ~10 ns and maintenance of a compact, well-behaved complex. (G) Free-energy landscape of the SIRT1–naringenin trajectory plotted along PC1 and PC2 (Δ G = − k B T ln P ), showing a dominant low-energy basin corresponding to the bound state. (H) SPR analysis of SIRT1–naringenin binding, showing concentration-dependent sensorgrams and a 1:1 Langmuir fit consistent with specific interaction. (I) Western blots of FOXO3 and Depp1 (HT22 and MC3T3-E1) after Sirt1 knockdown (KD) or overexpression (OE). (J) TMRE staining (ΔΨm) and (K) corresponding quantification: ΔΨm decreases with KD and increases with OE. (L) Triplex IF (LC3, TOM20, and DEPP1) showing the autophagy burden, mitochondrial network integrity, and DEPP1 levels across KD/OE conditions. (M) Quantifications of puncta burden, TOM20 continuity, and DEPP1 intensity. Docking to DEPP1 with neoeriocitrin, naringin, and naringenin produced low-affinity, nonconvergent poses (not shown); docking and molecular dynamics results for naringin, naringenin, and the positive control are provided in the Supplementary Materials. Abbreviations: R g , radius of gyration; RMSF, root mean square fluctuation; SASA, solvent-accessible surface area; RMSD, root mean square deviation.

Article Snippet: IF was performed on mouse hippocampal sections and on MC3T3-E1 and HT22 cells using primary antibodies DEPP1 (CUSABIO, CSB-PA865135LA01HU), LC3 (Immunoway, PT0235R), and TOM20 (Immunoway, PT0287R).

Techniques: Functional Assay, Solvent, Binding Assay, Concentration Assay, Western Blot, Knockdown, Over Expression, Staining, Produced, Positive Control

Journal: Research

Article Title: Targeting a Shared Mitophagy Regulator: The SIRT1–FOXO3–DEPP1 Axis Underpins the Dual Bone and Brain Benefits of Total Flavonoids from Drynaria fortunei

doi: 10.34133/research.1125

Figure Lengend Snippet: SIRT1 determines cellular responsiveness to TFDF in neurons and osteoblasts. Under H 2 O 2 injury, cells were treated with TFDF alone or in combination with Sirt1 knockdown (KD) or overexpression (OE). Western blots showed that TFDF decreased acetyl-FOXO3 and DEPP1 levels and that these effects were abrogated by KD and strengthened by OE in HT22 and MC3T3-E1 cells (A, densitometry in B). ROS imaging/quantification demonstrated that injury-induced oxidative stress was reduced by TFDF, partially reversed by KD, and further reduced by OE (C and D). Immunofluorescence staining for LC3B/TOM20/DEPP1 revealed a TFDF-driven improvement in autophagy–mitochondrial morphology that was attenuated by KD and potentiated by OE in both cell types (E and F). Autophagy immunoblots (LC3-II and p62) confirmed the same interaction pattern (G, densitometry in H).

Article Snippet: IF was performed on mouse hippocampal sections and on MC3T3-E1 and HT22 cells using primary antibodies DEPP1 (CUSABIO, CSB-PA865135LA01HU), LC3 (Immunoway, PT0235R), and TOM20 (Immunoway, PT0287R).

Techniques: Knockdown, Over Expression, Western Blot, Imaging, Immunofluorescence, Staining

Journal: Research

Article Title: Targeting a Shared Mitophagy Regulator: The SIRT1–FOXO3–DEPP1 Axis Underpins the Dual Bone and Brain Benefits of Total Flavonoids from Drynaria fortunei

doi: 10.34133/research.1125

Figure Lengend Snippet: TFDF alleviates OVX–CUMS-associated bone–brain comorbidity by engaging SIRT1-centered stress–autophagy signaling. Left: Experimental framework: 7-week-old female mice underwent ovariectomy combined with chronic unpredictable mild stress (OVX+CUMS, 12 weeks) and received total flavonoids of Drynaria fortunei (TFDF), resulting in improved bone density and depression-like behaviors. Right: Working model: OVX+CUMS-related cellular stress elevates reactive oxygen species (ROS) and disrupts autophagy–mitochondrial homeostasis. TFDF activates SIRT1, reduces FOXO3 acetylation, suppresses stress-responsive DEPP1, and restores antioxidant capacity (e.g., catalase [CAT]), thereby lowering ROS (with N-acetyl-L-cysteine [NAC], shown as an antioxidant control) and rebalancing autophagy to support organelle quality control. These coordinated effects ultimately improve cellular function and the observed functional phenotype in bone- and brain-relevant cells.

Article Snippet: IF was performed on mouse hippocampal sections and on MC3T3-E1 and HT22 cells using primary antibodies DEPP1 (CUSABIO, CSB-PA865135LA01HU), LC3 (Immunoway, PT0235R), and TOM20 (Immunoway, PT0287R).

Techniques: Control, Cell Function Assay, Functional Assay