Journal: Poultry science

Article Title: Duck Tembusu virus induced mitophagy in vacuolate spermatogenic cells is mediated by PINK1-Parkin pathway.

doi: 10.1016/j.psj.2025.104795

Figure Lengend Snippet: Fig. 2. The variation of spermatogenic cell after DTMUV infection. (A-D) Nucleus stained by DAPI. (E-H) Sertoli cells stained by SYCP3. (I-L) Merge pictures of DAPI and WT1 staining. Ctl: control group, 2 dpi: 2 days post-infection, 4 dpi: 4 days post-infection, 8 dpi: 8 days post-infection. Scar bars =20 μm.

Article Snippet: After incubation with SYCP3 Polyclonal antibody (1:200, 23024-1-AP, Proteintech, Wuhan) overnight at 4◦C, the sections were incubated via FITC-conjugated Goat anti-Rabbit IgG (H + L) (1:200, ABclonal, AS011, Wuhan) at 37◦C for 30 min.

Techniques: Infection, Staining, Control

Journal: PLOS ONE

Article Title: Short communication: Upregulation of hypoxia/reoxygenation-induced Shc3 by downregulated miR-455-5p, suppresses trophoblast invasion and is associated with placental inflammation and angiogenesis in preeclampsia

doi: 10.1371/journal.pone.0314544

Figure Lengend Snippet: (A) Heatmap of RNA-Seqs. (B) Selection of potential miR-455-5p downstream genes. (C) Schematic diagram of luciferase reporter-Shc3 3’UTR constructs. (D) The outcomes of miR-455-5p overexpression on the luciferase activity in HTR8/SVneo cells transfected with the WT or MUT luciferase reporter constructs. (E) The effect of miR-455-5p inhibition and overexpression on Shc3 mRNA expression in HTR8/SVneo cells. (F) The level of Shc3 protein expression after miR-455-5p inhibition and overexpression in HTR8/SVneo. Data presented shows the average values ± standard deviation (**p < 0.01, *p < 0.05). The findings presented were obtained from a minimum of three independent experiments.

Article Snippet: SHC3, Beta Actin antibodies and HRP-conjugated Affinipure Goat Anti Rabbit IgG(H+L) were obtained from Proteintech (Wuhan, China).

Techniques: Selection, Luciferase, Construct, Over Expression, Activity Assay, Transfection, Inhibition, Expressing, Standard Deviation

Journal: PLOS ONE

Article Title: Short communication: Upregulation of hypoxia/reoxygenation-induced Shc3 by downregulated miR-455-5p, suppresses trophoblast invasion and is associated with placental inflammation and angiogenesis in preeclampsia

doi: 10.1371/journal.pone.0314544

Figure Lengend Snippet: (A) The migration and invasion of HTR8/SVneo cells with miR-455-5p overexpression plasmids co-cultured with the plasmids of Shc3, or Shc3-NC in H/R. (B) Flow cytometry data of the apoptosis of HTR8/SVneo cells with miR-455-5p overexpression plasmids co-cultured with the plasmids expressing Shc3 or Shc3-NC in H/R. (C) Western blot results of Shc3 expression in HTR8/SVneo cells transfected with the miR-455-5p mimic, Shc3, or Shc3-NC. (D) The results of Shc3 expression examined by immunohistochemistry. Data presented indicate the average values ± standard deviation (**p < 0.01, *p < 0.05). The findings presented were obtained from a minimum of three independent experiments.

Article Snippet: SHC3, Beta Actin antibodies and HRP-conjugated Affinipure Goat Anti Rabbit IgG(H+L) were obtained from Proteintech (Wuhan, China).

Techniques: Migration, Over Expression, Cell Culture, Flow Cytometry, Expressing, Western Blot, Transfection, Immunohistochemistry, Standard Deviation

Journal: PLOS ONE

Article Title: Short communication: Upregulation of hypoxia/reoxygenation-induced Shc3 by downregulated miR-455-5p, suppresses trophoblast invasion and is associated with placental inflammation and angiogenesis in preeclampsia

doi: 10.1371/journal.pone.0314544

Figure Lengend Snippet: (A~B). H/R promotes the production of Shc3, which may be transported to the extracellular space through medium/large extracellular vesicles. (A) Western blot results of Shc3 expression in hypoxia, H/R, and normoxia, respectively. (B) Volcano plot of Shc3 mRNA expression in placental tissue, extracellular medium/large vesicles and small vesicles. (C~E). Shc3 is involved in placental inflammation and angiogenesis inhibition. (C) Hematoxylin-eosin staining results on normotensive placental tissues and high Shc3 expression PE placental tissues. (D) The level of Shc3 in EA.hy926 after transfection with the plasmids of Shc3, and Shc3-NC, in H/R compared with controls. (E) Angiogenesis of EA.hy926 after transfection with the plasmids of Shc3, and Shc3-NC in H/R, as determined by the number of master junction and master segment length.

Article Snippet: SHC3, Beta Actin antibodies and HRP-conjugated Affinipure Goat Anti Rabbit IgG(H+L) were obtained from Proteintech (Wuhan, China).

Techniques: Western Blot, Expressing, Inhibition, Staining, Transfection

Journal: Cell Discovery

Article Title: Luminal hormone-responsive cells tune the regenerative remodeling of mammary glands in large mammals

doi: 10.1038/s41421-025-00848-3

Figure Lengend Snippet: a UMAP plot displaying 14 identified cell types within the goat mammary glands. Cells are annotated and colored by type. Subsets of cell types including luminal, basal, fibroblast, immune, endothelial cell are labeled. b The percentages of −4W (red) and +1 W (blue) cells in each cell type are shown in a UMAP plot. c UMAP plots showing the expression of selected marker genes in four luminal subtypes. d Pseudotemporal trajectory analysis of scRNA-seq data of luminal secretory cells is shown in a UMAP plot. e Changes in the proportion of LumSecP and LumSec cells in luminal cell types were identified by scRNA-seq data at −4W and +1 W. n = 3 goats per group. f Representative images of tissue immunofluorescence staining for ALDH1A3 (red), KRT18 (green), and DAPI at −4W and +1 W. Scale bars, 50 μm. g Bar plots exhibiting the percentage of ALDH1A3-positive cells in luminal cells (labeled by KRT18) in f . n = 15 sections per group. h Representative images of tissue immunofluorescence staining for FABP3 (red), KRT18 (green), and DAPI at −4W and +1 W. Scale bars, 50 μm. i Bar plots exhibiting the percentage of FABP3-positive cells in luminal cells (labeled by KRT18) in h . n = 20 sections per group. The data are presented as the mean ± SEM. The P values of two-sided Student’s t -tests are shown in e , g , i .

Article Snippet: For immunofluorescence staining, goat mammary tissues were stained with primary antibodies against ALDH1A3 (#25167-1-AP, Proteintech), FABP3 (#10676-1-AP, Proteintech), KRT17 (#17516-1-AP, Proteintech), KRT14 (#SC65-06, HUABIO), KRT18 (conjugated with iFluorTM 488, #SZ80-07, HUABIO), KRT8 (#10384-1-AP, ProteinTech), PR (#25871-1-AP, ProteinTech), ER (#21244-1-AP, ProteinTech), and PCNA (#ab29, Abcam).

Techniques: Labeling, Expressing, Marker, Immunofluorescence, Staining

Journal: Cell Discovery

Article Title: Luminal hormone-responsive cells tune the regenerative remodeling of mammary glands in large mammals

doi: 10.1038/s41421-025-00848-3

Figure Lengend Snippet: a Relative proportions of LumHR cells in total luminal cells identified by scRNA-seq data at −4W and +1 W. n = 3 goats per group. b , c Bar plots exhibiting the percentage of PGR - and ESR1 -positive cells within luminal cells in scRNA-seq data. n = 3 goats per group. d Representative images of immunofluorescence staining for PR (red), KRT18 (green), and DAPI (blue). Scale bars, 50 μm. e Bar plots exhibiting the percentage of PR-positive cells in luminal cells (labeled by KRT18) in ( d ). n = 5 goats per group. f Representative images of immunofluorescence staining for ER (red), KRT18 (green), and DAPI (blue). Scale bars, 50 μm. g Bar plots exhibiting the percentage of ER-positive cells in luminal cells (labeled by KRT18) in ( f ). n = 5 goats. h Violin plot showing the specific expression of PRLR in LumHR cells by scRNA-seq. i Heatmap displaying the transcriptional level of indicated genes related to milk protein and luminal differentiation in the goat mammary organoids ( n = 3 biological replicates) treated with or without prolactin and in the mammary tissue at −4W (non-lactation) and +1 W (lactation). n = 3 goats for tissues. j Proportions of luminal cell types in goat mammary organoids incubated with or without prolactin predicted by CIBERSORTx deconvolution. n = 3 biological replicates. k Representative images of immunofluorescence staining for ALDH1A3 (red), KRT18 (green), and DAPI in mammary organoids incubated with or without prolactin. Scale bars, 10 μm. l Bar plots exhibiting the percentage of ALDH1A3-positive cells in luminal cells (labeled by KRT18) in ( k ). n = 14 domes in the control group and n = 10 domes in the prolactin treated group. m Representative images of immunofluorescence staining for FABP3 (red), KRT18 (green) and DAPI in mammary organoids incubated with or without prolactin. Scale bars, 10 μm. n Bar plots exhibiting the percentage of FABP3-positive cells in luminal cells (labeled by KRT18) in ( m ). n = 6 domes in the control group and n = 7 domes in the prolactin-treated group. The data are presented as the mean ± SEM. The P values of two-sided Student’s t -tests are shown in ( a – c , e , g , j , l , n ).

Article Snippet: For immunofluorescence staining, goat mammary tissues were stained with primary antibodies against ALDH1A3 (#25167-1-AP, Proteintech), FABP3 (#10676-1-AP, Proteintech), KRT17 (#17516-1-AP, Proteintech), KRT14 (#SC65-06, HUABIO), KRT18 (conjugated with iFluorTM 488, #SZ80-07, HUABIO), KRT8 (#10384-1-AP, ProteinTech), PR (#25871-1-AP, ProteinTech), ER (#21244-1-AP, ProteinTech), and PCNA (#ab29, Abcam).

Techniques: Immunofluorescence, Staining, Labeling, Expressing, Incubation, Control

Journal: Cell Discovery

Article Title: Luminal hormone-responsive cells tune the regenerative remodeling of mammary glands in large mammals

doi: 10.1038/s41421-025-00848-3

Figure Lengend Snippet: a Schematic illustration of targeted ablation of LumHR cells using the prlr -promoter to drive expression of DTA. b Experimental setup used in AAV intraductally injected mammary gland of ROSA-DTA +/ − mice under RR. c Whole-mount staining with carmine alum of mammary glands from ROSA-DTA +/ − mice (lactation day 2) intraductally injected with AAV-pPrlr-Cre or AAV-Control . Scale bars, 4 mm (top) and 500 μm (bottom). d , e Immunohistochemical staining and quantification of ER-positive luminal cells in mammary glands intraductally injected with AAV-pPrlr-Cre or AAV-Control. n = 3 mice per group. Scale bars, 50 μm. f , g Immunohistochemical staining and quantification of PR-positive luminal cells in mammary glands intraductally injected with AAV-pPrlr-Cre or AAV-Control . n = 3 mice per group. Scale bars, 50 μm. h , i Immunohistochemical staining and quantification of β-casein-positive alveoli number per mm 2 in mammary glands intraductally injected with AAV-pPrlr-Cre or AAV-Control . n = 3 mice per group. Scale bars, 50 μm. j Representative images of immunofluorescence staining for FABP3 (red), KRT18 (green), and DAPI in mammary glands intraductally injected with AAV-pPrlr-Cre or AAV-Control . Scale bars, 50 μm. k Bar plots exhibiting the percentage of FABP3-positive cells in luminal cells (labeled by KRT18) in j . n = 3 mice per group. l Representative images of immunofluorescence staining for ALDH1A3 (red), KRT18 (green), and DAPI in mammary glands intraductally injected with AAV-pPrlr-Cre or AAV-Contro l. Scale bars, 50 μm. m Bar plots exhibiting the percentage of FABP3-positive cells in luminal cells (labeled by KRT18) in ( l ). n = 3 mice per group. The data are presented as the mean ± SEM. The P values of two-sided Student’s t -tests are shown in d , g , i , l , m .

Article Snippet: For immunofluorescence staining, goat mammary tissues were stained with primary antibodies against ALDH1A3 (#25167-1-AP, Proteintech), FABP3 (#10676-1-AP, Proteintech), KRT17 (#17516-1-AP, Proteintech), KRT14 (#SC65-06, HUABIO), KRT18 (conjugated with iFluorTM 488, #SZ80-07, HUABIO), KRT8 (#10384-1-AP, ProteinTech), PR (#25871-1-AP, ProteinTech), ER (#21244-1-AP, ProteinTech), and PCNA (#ab29, Abcam).

Techniques: Expressing, Injection, Staining, Control, Immunohistochemical staining, Immunofluorescence, Labeling

Journal: OncoTargets and Therapy

Article Title:

Antitumor Effect of miR-1294/Pyruvate Kinase M2 Signaling Cascade in Osteosarcoma Cells

doi: 10.2147/ott.s232718

Figure Lengend Snippet: Figure 4 PKM2 was a target of miR-1294 in osteosarcoma cells. (A) PKM2 expression in human mesenchymal stem cells (hMSCs) and five osteosarcoma cell lines (Saos-2, MG63, U2OS, HOS, and 143B) was assessed by real-time PCR. (B) The miR-1294 binding site predictions for PKM2 3ʹUTR by microRNA.org. (C) A dual luciferase reporter assay was performed to confirm the binding in 293T cells. (D and E) PKM2 expression in U2OS and HOS cells transfected with miR-1294 mimic by real-time PCR and Western blot assays. (F and G) PKM2 expression in 143B cells transfected with miR-1294 inhibitor by real-time PCR and Western blot assays. Data are shown as mean ± SD. ∗P < 0.05, ∗∗∗P < 0.001.

Article Snippet: After dewaxing with xylene and rehydration with gradient ethanol, the sections were subjected to antigen retrieval and incubation with one of the primary antibodies against c-Myc (1:200; Proteintech), cyclin D1 (1:200; Boster), MMP2; (1:200; Proteintech), MMP-9 (1:200; Proteintech), and PKM2 (1:200; Proteintech) overnight at 4°C.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Binding Assay, Luciferase, Reporter Assay, Transfection, Western Blot

Journal: OncoTargets and Therapy

Article Title:

Antitumor Effect of miR-1294/Pyruvate Kinase M2 Signaling Cascade in Osteosarcoma Cells

doi: 10.2147/ott.s232718

Figure Lengend Snippet: Figure 5 Overexpression of PKM2 reversed the actions of miR-1294 upregulation on osteosarcoma cells. (A) Cell proliferation was evaluated by CCK-8 assay. (B) Cell apoptosis was examined by flow cytometry using Annexin Vand PI staining. (C) Cell migration was assessed by a wound-healing assay. (D) Cell invasiveness was determined by transwell assay. (E) Expression levels of c-Myc, cyclin D1, cleaved-caspase 3, MMP-2, and MMP-9 were texted by Western blotting. Data are shown as mean ± SD. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: After dewaxing with xylene and rehydration with gradient ethanol, the sections were subjected to antigen retrieval and incubation with one of the primary antibodies against c-Myc (1:200; Proteintech), cyclin D1 (1:200; Boster), MMP2; (1:200; Proteintech), MMP-9 (1:200; Proteintech), and PKM2 (1:200; Proteintech) overnight at 4°C.

Techniques: Over Expression, CCK-8 Assay, Cytometry, Staining, Migration, Wound Healing Assay, Transwell Assay, Expressing, Western Blot

Journal: OncoTargets and Therapy

Article Title:

Antitumor Effect of miR-1294/Pyruvate Kinase M2 Signaling Cascade in Osteosarcoma Cells

doi: 10.2147/ott.s232718

Figure Lengend Snippet: Figure 6 Overexpression of miR-1294 inhibited tumorigenicity of osteosarcoma cells. (A, B) Tumor volume in nude mice after subcutaneous injection of ME63 and U2O2 cells with or without overexpression of miR-1294. (C) The expression of miR-1294 in tumor tissues was determined by real-time PCR. (D, E) The expression of PKM2, c-Myc, cyclin D1, MMP-2, and MMP-9 in tumor tissues was assessed using immunohistochemistry and Western blot analysis. Data are shown as mean ± SD from 6 mice/ group. ∗P < 0.05, ∗∗∗P < 0.001.

Article Snippet: After dewaxing with xylene and rehydration with gradient ethanol, the sections were subjected to antigen retrieval and incubation with one of the primary antibodies against c-Myc (1:200; Proteintech), cyclin D1 (1:200; Boster), MMP2; (1:200; Proteintech), MMP-9 (1:200; Proteintech), and PKM2 (1:200; Proteintech) overnight at 4°C.

Techniques: Over Expression, Injection, Expressing, Real-time Polymerase Chain Reaction, Immunohistochemistry, Western Blot