p16  (Proteintech)

Journal: Materials Today Bio

Article Title: 3D-printed titanium scaffolds coated with a multifunctional photothermal-responsive hydrogel promote osteoporotic bone defect repair

doi: 10.1016/j.mtbio.2026.102879

Figure Lengend Snippet: Effects of hydrogel coatings on delaying senescence of BMSCs (A) Representative image of SA- β -gal staining. (B) Quantitative analysis of SA- β -gal staining intensity. (C) mRNA expression levels of p16 , p53 , Ccl2 , Tnf-α , IL-1β, and IL-6 in BMSCs. (D) Quantitative analysis of p21 fluorescence intensity. (E) Representative fluorescence images of p62 staining. (F) Quantitative analysis of p62 fluorescence intensity. (G) Representative fluorescence images of LC3B staining. (H) Quantitative analysis of LC3B fluorescence intensity. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001, n = 3.

Article Snippet: Primary antibodies against OPN (22952-1-AP), RUNX2 (20700-1-AP), CD86 (13395-1-AP), CD206 (18704–1-AP), p21 (10355-1-AP), p16 (10883-1-AP), LC3B (14600-1-AP), and CL594-phalloidin (PF00003) were obtained from Proteintech (Wuhan, China).

Techniques: Staining, Expressing, Fluorescence

Journal: bioRxiv

Article Title: The HSP90–CDC37 Chaperone System Orchestrates RAF1 Kinase Activation Through a Pre-Dimerization Mechanism

doi: 10.64898/2026.03.25.713956

Figure Lengend Snippet: A) Western blot analysis showing activation of the MAPK pathway in RAF-deficient MEFs. RAFlox MEFs were transduced in parallel with retroviruses expressing the indicated RAF isoforms (ARAF, BRAF, BRAF D594A , or BRAF V600E ), either alone (homodimers) or in combination with lentiviruses expressing RAF1 (heterodimers). Subsequently, cells were infected with adenoviruses encoding CRE recombinase to induce the deletion of all endogenous RAF genes. Phosphorylated MEK1/2 (pMEK1/2) levels were assessed by immunoblotting and quantified relative to parental RAFless cells, after normalization to the loading control. B) Colony formation assay of RAFless MEFs expressing the indicated RAF isoforms, with or without a shRNA targeting p16 INK4a to prevent cellular senescence. Ablation of the endogenous RAF genes was achieved as described in panel (a). Representative crystal violet–stained plates from one experiment are shown.

Article Snippet: Following blocking with 5% BSA in TBS-T for 1 hour at RT, the membranes were incubated with the following primary antibodies: anti-HSP90 (Santa Cruz, sc-13119; 1:1,000), anti-RAF1 (BD, 610151; 1:1,000), anti-CDC37 (Santa Cruz, sc-13129; 1:1,000), anti-pan-Ras (Calbiochem, OP40; 1:1,000), anti-V5-tag (ThermoFisher Scientific, R960-25; 1:1,000), anti-StrepTagII (ThermoFisher Scientific, MAS-37747; 1:500), anti-Vinculin (Sigma, V9131; 1:5,000), anti-BRAF (Santa Cruz, sc-5284; 1:1,000), anti-ARAF (Cell Signaling, 4432; 1:500), anti-phospho-MEK1/2 (Ser217/221) (Cell Signaling, 9121; 1:250), anti-MEK1 (Santa Cruz, sc-6250; 1:1,000), anti-ERK1/2 (BD, 554100/610103; 1:1,000), anti-phospho-ERK1/2 (Cell Signaling, 9101; 1:250) and anti-CDKN2A/p16-INK4A (Proteintech, 10883-1-AP; 1:500).

Techniques: Western Blot, Activation Assay, Expressing, Infection, Control, Colony Assay, shRNA, Staining

Journal: bioRxiv

Article Title: Circulating extracellular vesicles drive microglial senescence and neurodegeneration in Parkinson’s disease

doi: 10.64898/2026.03.10.709299

Figure Lengend Snippet: (a) PVDF membrane blotted with anti-APO B100, anti-TSG101, anti-Alix, anti-Calnexin, and anti-CD81 antibodies. Black boxes represent the cropped images shown in . (b-c) PVDF membranes blotted with anti-p16, anti-p21, and anti-β-actin antibodies. The black box represents the cropped images shown in .

Article Snippet: Membranes were blocked using Intercept (TBS) blocking buffer for 1 hour and then incubated overnight at 4°C with the following primary antibodies: anti-CD81 (1:500, Abcam, ab109201), anti-TSG101 (1:1000, Abcam, ab83), anti-ApoB (1:500, Abcam, ab20737), anti-Calnexin (1:500, Abcam, ab133615), anti-Alix (1:500, Abcam, ab186429), anti-p16 (1:1000, Proteintech, 10883-1-AP), anti-p21 (1:1000, Proteintech, 10355-1-AP), and anti-β-Actin (1:1000, SIGMA, A1978), overnight at 4°C.

Techniques: Membrane

Journal: bioRxiv

Article Title: Circulating extracellular vesicles drive microglial senescence and neurodegeneration in Parkinson’s disease

doi: 10.64898/2026.03.10.709299

Figure Lengend Snippet: (a) Immunostaining of HMC3 cells with DAPI (blue) and IBA1 (red) after 24 hours of EV treatment. Scale bar: 100 μm; zoom scale bar: 50 μm (b-c). Percentage and surface area of activated HMC3 cells after 24 hours of EV treatment. Data are represented as mean ± SEM and median with interquartile range (n=8 images per group). P-values are represented in the figures. (d) Heatmap showing normalized signal intensity of cytokine expression in HMC3 cells treated with PD-EV- and HC-EV . (e) Western blot quantification of p16 INK4a and p21 WAF /CIP in HMC3 cells after 48 and 96 hours of EV treatment (n=3). (f-g) Semi-quantification of p16 INK 4a and p21 WAF /CIP expressed as percentage change: (Normalized signal (EV-treated))/ (Normalized signal (PBS control)-1) *100. Data are represented as mean ± SEM. P-value is represented in the figures.

Article Snippet: Membranes were blocked using Intercept (TBS) blocking buffer for 1 hour and then incubated overnight at 4°C with the following primary antibodies: anti-CD81 (1:500, Abcam, ab109201), anti-TSG101 (1:1000, Abcam, ab83), anti-ApoB (1:500, Abcam, ab20737), anti-Calnexin (1:500, Abcam, ab133615), anti-Alix (1:500, Abcam, ab186429), anti-p16 (1:1000, Proteintech, 10883-1-AP), anti-p21 (1:1000, Proteintech, 10355-1-AP), and anti-β-Actin (1:1000, SIGMA, A1978), overnight at 4°C.

Techniques: Immunostaining, Expressing, Western Blot, Control

Journal: bioRxiv

Article Title: Circulating extracellular vesicles drive microglial senescence and neurodegeneration in Parkinson’s disease

doi: 10.64898/2026.03.10.709299

Figure Lengend Snippet: (a-c) p16 INK4a, p21 WAF /CIP , and SERPINE1 mRNA relative expression in HMC3 cells after 48 and 96 hours of treatment with HC-EV, PD-EV, or PBS.

Article Snippet: Membranes were blocked using Intercept (TBS) blocking buffer for 1 hour and then incubated overnight at 4°C with the following primary antibodies: anti-CD81 (1:500, Abcam, ab109201), anti-TSG101 (1:1000, Abcam, ab83), anti-ApoB (1:500, Abcam, ab20737), anti-Calnexin (1:500, Abcam, ab133615), anti-Alix (1:500, Abcam, ab186429), anti-p16 (1:1000, Proteintech, 10883-1-AP), anti-p21 (1:1000, Proteintech, 10355-1-AP), and anti-β-Actin (1:1000, SIGMA, A1978), overnight at 4°C.

Techniques: Expressing

Journal: bioRxiv

Article Title: Circulating extracellular vesicles drive microglial senescence and neurodegeneration in Parkinson’s disease

doi: 10.64898/2026.03.10.709299

Figure Lengend Snippet: (a) PVDF membrane blotted with anti-APO B100, anti-TSG101, anti-Alix, anti-Calnexin, and anti-CD81 antibodies. Black boxes represent the cropped images shown in . (b-c) PVDF membranes blotted with anti-p16, anti-p21, and anti-β-actin antibodies. The black box represents the cropped images shown in .

Article Snippet: Membranes were blocked using Intercept (TBS) blocking buffer for 1 hour and then incubated overnight at 4°C with the following primary antibodies: anti-CD81 (1:500, Abcam, ab109201), anti-TSG101 (1:1000, Abcam, ab83), anti-ApoB (1:500, Abcam, ab20737), anti-Calnexin (1:500, Abcam, ab133615), anti-Alix (1:500, Abcam, ab186429), anti-p16 (1:1000, Proteintech, 10883-1-AP), anti-p21 (1:1000, Proteintech, 10355-1-AP), and anti-β-Actin (1:1000, SIGMA, A1978), overnight at 4°C.

Techniques: Membrane

Journal: bioRxiv

Article Title: Circulating extracellular vesicles drive microglial senescence and neurodegeneration in Parkinson’s disease

doi: 10.64898/2026.03.10.709299

Figure Lengend Snippet: (a) Immunostaining of HMC3 cells with DAPI (blue) and IBA1 (red) after 24 hours of EV treatment. Scale bar: 100 μm; zoom scale bar: 50 μm (b-c). Percentage and surface area of activated HMC3 cells after 24 hours of EV treatment. Data are represented as mean ± SEM and median with interquartile range (n=8 images per group). P-values are represented in the figures. (d) Heatmap showing normalized signal intensity of cytokine expression in HMC3 cells treated with PD-EV- and HC-EV . (e) Western blot quantification of p16 INK4a and p21 WAF /CIP in HMC3 cells after 48 and 96 hours of EV treatment (n=3). (f-g) Semi-quantification of p16 INK 4a and p21 WAF /CIP expressed as percentage change: (Normalized signal (EV-treated))/ (Normalized signal (PBS control)-1) *100. Data are represented as mean ± SEM. P-value is represented in the figures.

Article Snippet: Membranes were blocked using Intercept (TBS) blocking buffer for 1 hour and then incubated overnight at 4°C with the following primary antibodies: anti-CD81 (1:500, Abcam, ab109201), anti-TSG101 (1:1000, Abcam, ab83), anti-ApoB (1:500, Abcam, ab20737), anti-Calnexin (1:500, Abcam, ab133615), anti-Alix (1:500, Abcam, ab186429), anti-p16 (1:1000, Proteintech, 10883-1-AP), anti-p21 (1:1000, Proteintech, 10355-1-AP), and anti-β-Actin (1:1000, SIGMA, A1978), overnight at 4°C.

Techniques: Immunostaining, Expressing, Western Blot, Control

Journal: bioRxiv

Article Title: Circulating extracellular vesicles drive microglial senescence and neurodegeneration in Parkinson’s disease

doi: 10.64898/2026.03.10.709299

Figure Lengend Snippet: (a-c) p16 INK4a, p21 WAF /CIP , and SERPINE1 mRNA relative expression in HMC3 cells after 48 and 96 hours of treatment with HC-EV, PD-EV, or PBS.

Article Snippet: Membranes were blocked using Intercept (TBS) blocking buffer for 1 hour and then incubated overnight at 4°C with the following primary antibodies: anti-CD81 (1:500, Abcam, ab109201), anti-TSG101 (1:1000, Abcam, ab83), anti-ApoB (1:500, Abcam, ab20737), anti-Calnexin (1:500, Abcam, ab133615), anti-Alix (1:500, Abcam, ab186429), anti-p16 (1:1000, Proteintech, 10883-1-AP), anti-p21 (1:1000, Proteintech, 10355-1-AP), and anti-β-Actin (1:1000, SIGMA, A1978), overnight at 4°C.

Techniques: Expressing