Journal: Frontiers in Physiology

Article Title: Mitophagy-associated biomarkers and macrophage involvement in pulmonary arterial hypertension: identification and functional implications

doi: 10.3389/fphys.2025.1673181

Figure Lengend Snippet: Identification of key cell type, pseudotime trajectories, and dynamic biomarker expression. (A) bar plot showing differential proportions of cell clusters between PAH and control samples; (B) Bubble plot illustrating the expression levels of the five candidate genes across annotated cell types; bubble size indicates the percentage of expressing cells, and colour intensity reflects expression magnitude; (C) heatmap of pathway enrichment analysis in the identified key cell population; (D–F) Pseudotime trajectory analysis of the key cell type: each point represents an individual cell coloured by pseudo time (dark blue = early state; light blue = late state). Black circles with numbers denote distinct cell‐state nodes identified during trajectory inference; (G, H) dynamic expression changes of BECN1, HIF1A, MFN1, RRAS, and TAX1BP1 along pseudotime in macrophage subtypes, stratified by cell state; (G) and macrophage polarization (M1 vs. M2) (H) ; (I) Violin plots showing state-specific expression patterns of the five hub genes.

Article Snippet: Membranes were incubated overnight at 4 °C with primary antibodies diluted in blocking buffer: RRAS (Proteintech, 66959-1-Ig; 1:1000), BECN1 (Proteintech, 66665-1-Ig; 1:1000), MFN1 (Zenbio, R27027 ; 1:1000), TAX1BP1 (Hanan Biotechnology, HA721648; 1:1000), HIF1A (Abcam, ab179483; 1:1000), and β-actin (Proteintech, 66009-1-Ig; 1:25,000).

Techniques: Biomarker Discovery, Expressing, Control

Journal: Frontiers in Physiology

Article Title: Mitophagy-associated biomarkers and macrophage involvement in pulmonary arterial hypertension: identification and functional implications

doi: 10.3389/fphys.2025.1673181

Figure Lengend Snippet: Validation of the Relationship between Biomarkers and Mitochondria. (A) expression levels of RRAS, BECN1, MFN1, HIF1A and TAX1BP1 in rat lung tissues detected by qPCR; (B) protein expression levels of RRAS, BECN1, MFN1, HIF1A and TAX1BP1 in rat lung tissues detected by WB; (C) expression and localization of LC3 and iNOS detected by immunofluorescence double-labeling colocalization analysis; (D) relative expression levels of iNOS in lung tissues; (E) relative expression levels of autophagy marker LC3 in lung tissues; (F) relative colocalization levels of LC3 and iNOS in lung tissues of CK and PAH; (G) normalized Pearson’s correlation coefficient (Pearson’s R) for colocalization of LC3 + iNOS in lung tissues Scale bar 100 μm; (H) Schematic diagram of the relationship between the five biomarkers and mitochondria. In the figures, ns indicates p > 0.05; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

Article Snippet: Membranes were incubated overnight at 4 °C with primary antibodies diluted in blocking buffer: RRAS (Proteintech, 66959-1-Ig; 1:1000), BECN1 (Proteintech, 66665-1-Ig; 1:1000), MFN1 (Zenbio, R27027 ; 1:1000), TAX1BP1 (Hanan Biotechnology, HA721648; 1:1000), HIF1A (Abcam, ab179483; 1:1000), and β-actin (Proteintech, 66009-1-Ig; 1:25,000).

Techniques: Biomarker Discovery, Expressing, Immunofluorescence, Labeling, Marker

Journal: bioRxiv

Article Title: Mechanosensitive ion channel PIEZO1 enhances endometrial decidualization through BECN1-dependent autophagy

doi: 10.1101/2025.08.05.668790

Figure Lengend Snippet: (A) Expression of LC3A/B in artificial decidualized uteri of Piezo1 f/f ( P1 f/f ) and Piezo1 d/d ( P1 d/d ) mice 2 or 4 days after stimulation; (B) Representative TEM images of 2-day in vitro decidualized mESCs (E 2 +P 4 treatment, EP) isolated from Piezo1 f/f ( P1 f/f ) and Piezo1 d/d ( P1 d/d ) mice, vehicle was used as control (CON), red arrows represent autophagosomes; (C) The quantification of autophagosomes in B; (D) The number of autophagosomes and autolysosomes in 2-day in vitro decidualized mESCs (E 2 +P 4 treatment, EP) isolated from Piezo1 f/f ( P1 f/f ) and Piezo1 d/d ( P1 d/d ) mice, detected by LC3-GFP-RFP vector, representative images shown in figure S4A; (E) Expression of LC3A/B in 2-day in vitro decidualized mESCs (E 2 +P 4 treatment, EP) co-treated with Yoda1 (Y), BAPTA-AM (B), KN93 (K), or U0126 (U); (F) Expression of LC3A/B in 2-day in vitro decidualized HESCs (MPA+cAMP treatment, Mc) co-treated with Yoda1 (Y), BAPTA-AM (B), KN93 (K), or U0126 (U); (G) Expression of p-BECN1 in artificial decidualized uteri of Piezo1 f/f ( P1 f/f ) and Piezo1 d/d ( P1 d/d ) mice 2 or 4 days after stimulation; (H) Expression of p-BECN1 in 2-day in vitro decidualized mESCs (E 2 +P 4 treatment, EP) co-treated with Yoda1 (Y), BAPTA-AM (B), KN93 (K), or U0126 (U); (I) Expression of p-BECN1 in 2-day in vitro decidualized HESCs (MPA+cAMP treatment, Mc) co-treated with Yoda1 (Y), BAPTA-AM (B), KN93 (K), or U0126 (U); (J) mRNA expression of decidualization marker Prl8a2 in 2-day in vitro decidualized, scrambled (siScra) or Beclin1 siRNA (siBeclin1) transfected mESCs (E 2 +P 4 treatment) isolated from Piezo1 f/f ( P1 f/f ) and Piezo1 d/d ( P1 d/d ) mice, co-cultured with or without Yoda1; (K) mRNA expression of decidualization marker Prl8a2 in 2-day in vitro decidualized mESCs (E 2 +P 4 treatment) isolated from Piezo1 f/f ( P1 f/f ) and Piezo1 d/d ( P1 d/d ) mice, co-cultured with different dose of Tat-BECLIN1; (L) mRNA expression of decidualization marker genes in 2-day in vitro decidualized HESCs (MPA+cAMP treatment), co-cultured with different doses of Tat-BECLIN1; Scale bar = 500 nm; ns p>0.05; *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001.

Article Snippet: The primary antibodies used in this study included phosphorylated CaMKII (pCaMKII, 1:1000, af3493, Affinit), phosphorylated ERK1/2 (p-ERK1/2, 1:1000, 4370, Cell Signaling Technology), total ERK1/2 (t-ERK1/2, 1:1000, 4695, Cell Signaling Technology), Cyclin D1 (1:1000, 2978, Cell Signaling Technology), CDK4 (1:1000, 23972, Cell Signaling Technology), LC3A/B (1:1000, 4108, Cell Signaling Technology), phosphorylated BECN1 (p-BECN1, 1:1000, PA5-112018, Thermo Fisher Scientific), total BECN1 (t-BECN1, 1:1000, 3495s, Cell Signaling Technology), phosphorylated mTOR (p-mTOR, 1:1000, 2971, Cell Signaling Technology), TUBULIN (1:1000, 2144 S, Cell Signaling Technology), and GAPDH (1:1000, sc-32233, Santa Cruz Biotechnology).

Techniques: Expressing, In Vitro, Isolation, Control, Plasmid Preparation, Marker, Transfection, Cell Culture

Journal: Materials Today Bio

Article Title: A biomimetic multimodal nanoplatform combining neutrophil-coated two-dimensional metalloporphyrinic framework nanosheet and exendin-4 to treat obesity-related osteoporosis

doi: 10.1016/j.mtbio.2025.102009

Figure Lengend Snippet: TET2 enhances mitophagy in DPA@NM@CTZE. ( A-B ) WB analysis assessed changes in TET2 and autophagosome proteins BCL2 and BECN1, with data shown as Mean ± SD. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, n = 3.

Article Snippet: WB using antibodies including: TET2 (Abcam, USA), COL1A1 (Collagen I, Abcam, UK), RUNX2 (Runt-related transcription factor 2, CST, USA), PINK1 (PTEN-induced putative kinase protein 1, Bioss, China), Parkin (E3 ubiquitin-protein ligase parkin, Servicebio, China), LC3B (Light Chain 3, Abmart, China), ATG4B (Autophagy Related 4B Cysteine Peptidase, Proteintech, China), BCL2 (B-cell lymphoma-2, Abmart, China), BECN1 (Beclin-1, Abmart, China), ACTB (Beyotime, China), Integrin β1 (MCE, USA), Integrin β2 (Bio-Techne, USA), CXCR2 (Proteintech, China), HRP-labeled Goat Anti-Rabbit IgG (H + L) (Beyotime, China), HRP-labeled Goat Anti-Mouse IgG (H + L) (Beyotime, China), KEAP1 (Kelch Like ECH Associated Protein 1, Proteintech, China),NRF2 (Nuclear factor erythroid 2-related factor 2, Proteintech, China).

Techniques: