Journal: Translational Psychiatry

Article Title: Double-hit of MIA and Nod2 deficiency induces sex-specific offspring behavioral abnormalities through placental dysregulation

doi: 10.1038/s41398-025-03747-z

Figure Lengend Snippet: A Hematoxylin and eosin staining of placental tissue. Left: Overall structure (Scale bar: 1000 μm); Right: Magnification of sinusoids (Scale bar: 100 μm). B Immunofluorescence for cluster of differentiation 31 ( CD31 ) (red) and 4′,6-diamidino-2-phenylindole (DAPI) (blue). From left to right: Overview (Scale bar: 1000 μm), CD31 channel, DAPI channel, and merged image (Scale bar: 100 μm). C Immunofluorescence for cytokeratin 7 ( CK7 ) (red) and DAPI (blue). Panels arranged as in B. D Comparison of placental efficiency (fetal-to-placental weight ratio) between groups. E Ratio of labyrinth zone to total area (labyrinth + junction zone). F Quantification of CD31 fluorescence intensity. G Quantification of CK7 fluorescence intensity. Sample size: n = 12 per group (male: female = 1:1). Data expressed as mean ± SEM; Student’s t -test or the non-parametric Wilcoxon rank-sum test.

Article Snippet: Subsequently, the sections were incubated separately with primary antibodies against cytokeratin 7 ( CK7 ) (1:400, 17513-1-AP; Proteintech) and cluster of differentiation 31 ( CD31 ) (1:400, 28083-1-AP; Proteintech), each diluted in PBS, overnight at 4 °C.

Techniques: Staining, Immunofluorescence, Comparison, Fluorescence

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: p16INK4a downregulation alleviates temporomandibular joint osteoarthritis combined with type 2 diabetes by driving M2 polarization.

doi: 10.1016/j.biopha.2025.118172

Figure Lengend Snippet: Fig. 3. MS37452 and P16KD restrain inflammatory angiogenesis in inflammatory microenvironment of TMJ. (A) A diagram showing how to collect conditioned medium (CM) from M0. M0 cells were divided into three groups: Ctrl, P16KD, and MS37452. M0 were stimulated with or without TNFα ± GP during culture for 24 h. Next, medium was changed to fresh RPMI-1640 + 10 % FBS for 24 h and M0 CM was collected. At last, M0 CM was co-cultured with HUVECs. Then, HUVECs were used for subsequent transwell assay, scratch assay, tube formation assay, western blot and RT-qPCR. (B) Representative images and quantification of HUVECs transwell assay (Ctrl: M0 VS TNFα+GP: t = 66.86, df = 4. Ctrl-TNFα+GP VS P16KD-TNFα+GP: t = 77.36, df = 4. Ctrl-TNFα+GP VS MS37452-TNFα+GP: t = 65.01, df = 4). (C) Representative images and quantification of HUVECs scratch assay (Ctrl: M0 VS TNFα+GP: t = 23.12, df = 4. Ctrl-TNFα+GP VS P16KD-TNFα+GP: t = 23.14, df = 4. Ctrl-TNFα+GP VS MS37452-TNFα+GP: t = 22.50, df = 4). (D) Representative images and quantification of HUVECs tube formation assay (Ctrl: M0 VS TNFα+GP: t = 139.4, df = 4. Ctrl-TNFα+GP VS P16KD-TNFα+GP: t = 115.5, df = 4. Ctrl-TNFα+GP VS MS37452-TNFα+GP: t = 116.8, df = 4). (E) Western blot analysis of CD31 and VEGFA in HUVECs from different groups. (F) RT-qPCR analysis of CD31 and VEGFA in HUVECs from different groups (Relative expression of CD31 mRNA: Ctrl: M0 VS TNFα+GP: t = 66.96, df = 4. Ctrl-TNFα+GP VS P16KD-TNFα+GP: t = 80.36, df = 4. Ctrl-TNFα+GP VS MS37452-TNFα+GP: t = 86.18, df = 4. Relative expression of VEGFA mRNA: Ctrl: M0 VS TNFα+GP: t = 61.40, df = 4. Ctrl-TNFα+GP VS P16KD-TNFα+GP: t = 75.08, df = 4. Ctrl-TNFα+GP VS MS37452-TNFα+GP: t = 72.31, df = 4). Each experiment was performed three to five times. The data are presented as mean ± SD. P16KD, p16 knockdown; Ctrl, control; M0, human THP-1 derived macrophages; GP, high glucose and high palmitoleic acid. *P < 0.05 represent significant differences between the indi cated columns.

Article Snippet: After that, membranes were incubated at 4 ◦C overnight with primary antibodies against inducible nitric oxide synthase (INOS) (Proteintech Group, Rosemont, IL, USA), arginase 1 (Arg1) (Proteintech Group, Rosemont, IL, USA), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Proteintech Group, Rosemont, IL, USA), transferrin receptor (TFRC) (Proteintech Group, Rosemont, IL, USA), vascular endothelial growth factor A (VEGFA) (Proteintech Group, Rosemont, IL, USA), alkaline phosphatase (ALP) (Proteintech Group, Rosemont, IL, USA), runt-related transcription factor 2 (RUNX2) (Proteintech Group, Rosemont, IL, USA), cluster differentiation 31 (CD31) (Proteintech Group, Rosemont, IL, USA), ferritin heavy chain (FTH1) (Proteintech Group, Rosemont, IL, USA).

Techniques: Cell Culture, Transwell Assay, Wound Healing Assay, Tube Formation Assay, Western Blot, Quantitative RT-PCR, Expressing, Knockdown, Control, Derivative Assay