antibodies hsp70  (Proteintech)

Journal: Bioactive Materials

Article Title: Near infrared enhanced palladium loaded siraitia grosvenorii carbon dots amplify mitophagy for acute lung injury immunotherapy

doi: 10.1016/j.bioactmat.2026.02.040

Figure Lengend Snippet: In vivo ALI therapy evaluation. A) Time schedule of in vivo animal experiment. B) Macroscopic observation in the lung tissue of treated rats. C) Wet/dry ratio in the lung tissue of treated rats. D) Inflammatory factors expression levels in the blood of treated rats. E) Inflammatory factors expression levels in the lung tissue of treated rats. F) ROS levels in the lung tissue of treated rats. (Scale bar = 50 μm) G) H&E staining images in the lung tissue of treated rats. (Scale bar = 100 μm) H) TNF-α expression levels in the lung tissue of treated rats, (Scale bar = 100 μm) and the corresponding quantified results (I). J) HSP70 expression levels in the lung tissue of treated rats, (Scale bar = 100 μm) and the corresponding quantified results (K). L) CD31 expression levels in the lung tissue of treated rats, (Scale bar = 100 μm) and the corresponding quantified results (M). The corresponding groups were: rats without treatments (sham group), and rats pretreated with LPS followed by IT administration of PBS (ALI group), CPs (CPs), CPs@SS31 (CPs@SS31) and CPs@SS31 combining with NIR irradiation (CPs@SS31+NIR). (”∗” symbol compared with sham group, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 and ∗∗∗∗p < 0.0001).

Article Snippet: And then, the cells were incubated with primary antibodies (anti-IL-6, TNF-α, CD206, CD86, CD31, HSP70 and PINK1, 1 : 200, Proteintech, USA) overnight.

Techniques: In Vivo, Expressing, Staining, Irradiation

Journal: Non-coding RNA Research

Article Title: CircEif3c/miR-96–5p/PHF20L1/MEOX2 axis in perivascular preadipocyte exosomes mediates fibroblast dysfunction and vascular remodeling

doi: 10.1016/j.ncrna.2026.01.006

Figure Lengend Snippet: PVPAC/AF co-culture model confirms that PVPAC-derived exosomes mediated intercellular communication. (A) Schematic diagram of primary PVPAC/AF cells culture with subsequent exosome isolation. (B) PVPAC/AF cells co-culture model. (B1) Schematic of the transwell-based co-culture setup. (B2) Representative TEM micrograph showing exosome morphology, scale bar = 100 nm. (B3) NTA-derived size distribution and concentration profiles of isolated exosomes. (B4) Crystal violet assay assessing cell proliferation under different glucose conditions, scale bar = 200 μm. (B5 and B6) Quantitative histograms corresponding to (B3) and (B4), respectively. Data are compared across mono-vs. co-culture systems under normal (NG) or high glucose (HG). vs NG + AF group, ∗P < 0.05, ∗∗P < 0.01. (C) Confocal microscopy tracking exosome uptake. Scale bar = 50 μm. (C1) PKH67-labeled PVPAC-derived exosomes (green) enriched in PVPAC cytoplasm. (C2) PKH67-labeled AF-derived exosomes abundant within AF cytoplasm. (C3) Time-course imaging displayed PVPAC-Exo accumulation in AFs, peaking at 4 h. (D) Quantification of migration and proliferation capacities in AFs after 24-h treatment with PVPAC-Exo (1 × 10 6 particles/mL), using PBS as a vehicle control, scale bar = 200 μm. (E) Impact of NG, HG, and GW4869 on exosome biology, scale bar = 100 nm. (E1) Morphology assessed by TEM. (E2) Proliferation measured via crystal violet. (E3) Western blot quantification of vimentin and exosomal markers (CD63, TSG101) in AFs. (F) RT-PCR analysis of circEif3c and miR-96–5p in AFs and PVPACs after 24 h NG vs. HG. HG induced highest circEif3c and lowest miR-96–5p expression in PVPACs. (G–K) Systematic comparison of exosomal protein signatures across culture modalities. (G1)Single-cell culture. (G2)Dual-cell co-culture. (G3) Co-culture pre-loaded with 1 × 10 6 /mL PVPAC-Exo. (H–K) Bar graphs present mean ± SD. n (the number of experiments) = 3; one-way ANOVA with Dunnett's post-test. ∗vs. respective NG group: ∗P < 0.05, ∗∗P < 0.01; vs. respective HG group: #P < 0.05, ##P < 0.01.

Article Snippet: Antibodies against MEOX2 (1:1500, #ab262916, Abcam, UK), PHF20L1 (1:1500, #ab118190, Abcam, UK), β-actin (#AC004, 1:5000, ABclone, Wuhan), Bcl-2 (1:2000, #ab182858, Abcam, UK), N-cadherin (1:1500, Abcam, UK), vimentin (1:2000, #ab92547, Abcam, UK), Anti-CD63 (1:1000, #ab315108, Abcam, UK), and TSG101 (1:2000, #28283-1-AP, Proteintech, USA) were purchased.

Techniques: Co-Culture Assay, Derivative Assay, Isolation, Concentration Assay, Crystal Violet Assay, Confocal Microscopy, Labeling, Imaging, Migration, Control, Western Blot, Reverse Transcription Polymerase Chain Reaction, Expressing, Comparison, Single Cell

Journal: Bioactive Materials

Article Title: A foam cell-targeted lipophagy restoration strategy stabilizes vulnerable atherosclerotic plaques

doi: 10.1016/j.bioactmat.2026.02.041

Figure Lengend Snippet: In vitro evaluation of foam cell lipid accumulation and lipophagy activation following OPN-HMCN@MLT treatment. ( A - C ) ORO and BODIPY staining images and corresponding quantification of ORO and BODIPY positive areas of RAW264.7 cells under different stimulations (n = 5, scale bar for ORO: 100 μm, scale bar for BODIPY: 20 μm). ( D ) Bio-TEM images of RAW264.7 cells post various treatments (n = 5, scale bars 1.0 μm). Green arrows indicate nanoparticles. ( E , F ) Morphometric analysis determined the mean number and area (μm 2 ) of LDs per cell section. ( G ) Confocal images depicting lipophagy flux in foam cells following different treatments (n = 5 biological replicates, scale bars: 10 μm). ( H - J ) The quantities of acidified autophagosomes (GFP-RFP+), neutral autophagosomes (GFP + RFP+), and LDs labeled with BODIPY were measured per cell for each condition. (K to N) Representative Western blot images and quantitative analysis of LC3, LAMP1, and P62 expression in foam cells. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001.

Article Snippet: To block nonspecific binding, membranes were incubated with 5% skim milk for 1 h. Thereafter, membranes were incubated overnight at 4 °C with primary antibodies against ABCA1, ABCG1, ACOX1, CPT1A, LC3 (ab192890, 1:2000, abcam), LAMP1 (84658-5-RR, 1:8000, Proteintech), PPARα (66826-1-Ig, 1:3000, Proteintech), PPARγ (66936-1-Ig, 1:10000, Proteintech), P62 (18420-1-AP, 1:10000, Proteintech), MCAD (55210-1-AP, 1:3000, Proteintech), LCAD (17526-1-AP, 1:10000, Proteintech), tubulin (80762-1-RR, 1:10000, Proteintech), GAPDH (60004-1-Ig, 1:50000, Proteintech), and β-actin (66009-1-Ig, 1:20000, Proteintech).

Techniques: In Vitro, Activation Assay, Staining, Labeling, Western Blot, Expressing

Journal: Bioactive Materials

Article Title: A foam cell-targeted lipophagy restoration strategy stabilizes vulnerable atherosclerotic plaques

doi: 10.1016/j.bioactmat.2026.02.041

Figure Lengend Snippet: Schematic of the anti-atherosclerotic mechanism of OPN-HMCN@MLT. ( A ) The study commenced with the synthesis of mesoporous carbon nanospheres (MCN) functionalized with an OPN-binding peptide and hyaluronic acid to construct the OPN-HMCN nanoplatform. The OPN-binding peptide was designed to recognize OPN enriched in the extracellular matrix and on the surface of foam cells, thereby enabling selective accumulation in OPN-rich pathological regions. Following OPN recognition, OPN-HMCN@MLT undergoes CD44-dependent endocytosis. Melatonin (MLT), a lipid autophagy–promoting agent, was subsequently encapsulated within the nanocarrier to form OPN-HMCN@MLT. Firstly, the released MLT can bind to and upregulate the expression of PPARα and PPARγ, which then promote the expression of downstream genes (ABCA1, ABCG1, ACOX-1, and CTP1A) and trigger the lipophagy. ( B ) Subsequently, its lipophagy-enhancing effects, including ABCA1/G1-mediated cholesterol efflux and CTP1A/ACOX-1-mediated mitochondrial fatty acid oxidation, were studied to confirm the reversal of foam cell formation. ( C ) These effects eventually promote foam cells to reverse into macrophages. Abbreviations: MCN, mesoporous carbon nanoparticle; OPN, osteopontin; MLT, melatonin; LDL, low-density lipoprotein; ox-LDL, oxidized low-density lipoprotein; PA, Photoacoustic.

Article Snippet: To block nonspecific binding, membranes were incubated with 5% skim milk for 1 h. Thereafter, membranes were incubated overnight at 4 °C with primary antibodies against ABCA1, ABCG1, ACOX1, CPT1A, LC3 (ab192890, 1:2000, abcam), LAMP1 (84658-5-RR, 1:8000, Proteintech), PPARα (66826-1-Ig, 1:3000, Proteintech), PPARγ (66936-1-Ig, 1:10000, Proteintech), P62 (18420-1-AP, 1:10000, Proteintech), MCAD (55210-1-AP, 1:3000, Proteintech), LCAD (17526-1-AP, 1:10000, Proteintech), tubulin (80762-1-RR, 1:10000, Proteintech), GAPDH (60004-1-Ig, 1:50000, Proteintech), and β-actin (66009-1-Ig, 1:20000, Proteintech).

Techniques: Binding Assay, Construct, Expressing