rabbit anti p16 ink4a polyclonal antibody  (Proteintech)

Journal: Frontiers in Pharmacology

Article Title: Targeted ErbB4 receptor activation prevents D-galactose-induced neuronal senescence via inhibiting ferroptosis pathway

doi: 10.3389/fphar.2025.1528604

Figure Lengend Snippet: Effect of ErbB4 receptor agonist and melatonin can ameliorate D-gal-induced aging in hippocampus in mice. (A) Western blot analysis of Lamin B1, p53, p21, p16, GFAP, and Iba-1 protein expression levels in the hippocampus of mice. (B–G) Quantification of Lamin B1, p53, p21, p16, GFAP, and Iba-1 protein levels. Data are presented as mean ± SD, with n = 4–5. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: The antibodies used, diluted in primary antibody diluents (P0023A, Beyotime), included: rabbit anti-phospho-ErbB4 (bs-3220R, Bioss; 1:1,000), rabbit anti-ErbB4 (AF6807, Beyotime; 1:1,000), rabbit anti-phospho-Akt1 (AF1546, Beyotime; 1:1,000), mouse anti-Akt1 (sc-377457, Santa Cruz Biotech; 1:1,000), rabbit anti-Nrf2 (AF7623, Beyotime; 1:1,000), rabbit anti-TFRC (AF8136, Beyotime; 1:1,000), rabbit anti-SLC7A11 (GB115276, Servicebio Biotech; 1:1,000), rabbit anti-GPX4 (AF7020, Beyotime; 1:1,000), rabbit anti-Lamin B1 (12987-1-AP, Proteintech, 1:5,000), rabbit anti-p53 (AF5258, Beyotime; 1:1,000), mouse anti-p21 (AP021, Beyotime; 1:1,000), rabbit anti-p16 (bs-23797R, Bioss Biotech, Beijing, China; 1:1,000), and rabbit anti-GAPDH (GB15002, Servicebio Biotech; 1:2000).

Techniques: Western Blot, Expressing

Journal: Frontiers in Pharmacology

Article Title: Targeted ErbB4 receptor activation prevents D-galactose-induced neuronal senescence via inhibiting ferroptosis pathway

doi: 10.3389/fphar.2025.1528604

Figure Lengend Snippet: The mitigation of D-gal-induced ferroptosis in HT22 cells by ErbB4 receptor agonist and melatonin. (A) Cell viability post D-gal exposure, evaluated using the CCK-8 assay (n = 5). (B) Cell viability following co-treatment with D-gal and ErbB4 receptor agonist, assessed via the CCK-8 assay (n = 6). (C) Cell viability following co-treatment with D-gal and melatonin, determined by the CCK-8 assay (n = 6). (D) Visualization of cellular senescence through SA-β-gal staining. Scale bar = 100 μm. (E) Quantification of SA-β-gal staining intensities (n = 3). (F) Western blot analysis was conducted to assess the expression levels of senescence-associated markers and ferroptosis-related proteins in HT22 cells. (G–M) Quantification of Nrf2, TFRC, SLC7A11, GPX4, p53, p21, and p16 levels was performed, with normalization to GAPDH (n = 4–7). (N) Intracellular GSH levels (n = 4). (O) Intracellular MDA levels (n = 5). (P) Quantitative analysis of ROS levels was undertaken (n = 4). (Q) Intracellular ROS generation was measured using DCFH-DA, wherein ROS activity was reflected as green fluorescence intensity. Scale bar = 200 μm. Data are presented as mean ± SD. Statistical significance is indicated as follows: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: The antibodies used, diluted in primary antibody diluents (P0023A, Beyotime), included: rabbit anti-phospho-ErbB4 (bs-3220R, Bioss; 1:1,000), rabbit anti-ErbB4 (AF6807, Beyotime; 1:1,000), rabbit anti-phospho-Akt1 (AF1546, Beyotime; 1:1,000), mouse anti-Akt1 (sc-377457, Santa Cruz Biotech; 1:1,000), rabbit anti-Nrf2 (AF7623, Beyotime; 1:1,000), rabbit anti-TFRC (AF8136, Beyotime; 1:1,000), rabbit anti-SLC7A11 (GB115276, Servicebio Biotech; 1:1,000), rabbit anti-GPX4 (AF7020, Beyotime; 1:1,000), rabbit anti-Lamin B1 (12987-1-AP, Proteintech, 1:5,000), rabbit anti-p53 (AF5258, Beyotime; 1:1,000), mouse anti-p21 (AP021, Beyotime; 1:1,000), rabbit anti-p16 (bs-23797R, Bioss Biotech, Beijing, China; 1:1,000), and rabbit anti-GAPDH (GB15002, Servicebio Biotech; 1:2000).

Techniques: CCK-8 Assay, Staining, Western Blot, Expressing, Activity Assay, Fluorescence

Journal: Frontiers in Pharmacology

Article Title: Targeted ErbB4 receptor activation prevents D-galactose-induced neuronal senescence via inhibiting ferroptosis pathway

doi: 10.3389/fphar.2025.1528604

Figure Lengend Snippet: The effects of ErbB4 receptor agonist and melatonin treatment on ferroptosis in Erastin-exposed HT22 cells. (A) Cell viability of HT22 cells post-Erastin exposure was evaluated using the CCK-8 assay (n = 5). (B) Cell viability was evaluated following treatment with Erastin, cotreatment with either E4A or melatonin, or both, utilizing the CCK-8 assay (n = 4). (C) Western blot analysis was conducted to determine the expression levels of senescence-associated markers and ferroptosis-related proteins in HT22 cells. (D–I) Quantitative analysis of Nrf2, TFRC, SLC7A11, GPX4, p21, and p16 levels, normalized to GAPDH (n = 4–7). (J) Intracellular ROS generation was measured using DCFH-DA, with ROS activity indicated by green fluorescence. Scale bar = 200 μm. (K) The quantitative analysis of ROS levels was conducted (n = 4). The data are presented as mean ± SD. Statistical significance is indicated as follows: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: The antibodies used, diluted in primary antibody diluents (P0023A, Beyotime), included: rabbit anti-phospho-ErbB4 (bs-3220R, Bioss; 1:1,000), rabbit anti-ErbB4 (AF6807, Beyotime; 1:1,000), rabbit anti-phospho-Akt1 (AF1546, Beyotime; 1:1,000), mouse anti-Akt1 (sc-377457, Santa Cruz Biotech; 1:1,000), rabbit anti-Nrf2 (AF7623, Beyotime; 1:1,000), rabbit anti-TFRC (AF8136, Beyotime; 1:1,000), rabbit anti-SLC7A11 (GB115276, Servicebio Biotech; 1:1,000), rabbit anti-GPX4 (AF7020, Beyotime; 1:1,000), rabbit anti-Lamin B1 (12987-1-AP, Proteintech, 1:5,000), rabbit anti-p53 (AF5258, Beyotime; 1:1,000), mouse anti-p21 (AP021, Beyotime; 1:1,000), rabbit anti-p16 (bs-23797R, Bioss Biotech, Beijing, China; 1:1,000), and rabbit anti-GAPDH (GB15002, Servicebio Biotech; 1:2000).

Techniques: CCK-8 Assay, Western Blot, Expressing, Activity Assay, Fluorescence

Journal: Frontiers in Pharmacology

Article Title: Targeted ErbB4 receptor activation prevents D-galactose-induced neuronal senescence via inhibiting ferroptosis pathway

doi: 10.3389/fphar.2025.1528604

Figure Lengend Snippet: Schematic diagram illustrating the effects of targeted activation of ErbB4 receptor on neuronal aging via ferroptosis inhibition. Small molecule agonist (E4A)can activate ErbB4 receptors and regulate the Akt/Nrf2 signaling pathway to achieve neuroprotective effect in D-gal-induced neuronal aging. D-gal treatment increases the accumulation of advanced glycation end products (AGEs) which stimulates the production of reactive oxygen species (ROS) and increase the expression of senescence marker genes P53, P16, and P21. E4A can promote Akt1 phosphorylation and promote Nrf2 entrance into the nucleus, in which it involves in regulating the expression of ferroptosis suppressor gene, such as SLC7A11 and GPX4 to attenuate lipid peroxidation, which can inhibit cell ferroptosis and neuronal aging. The diagram was created with MedPeer ( www.medpeer.cn ).

Article Snippet: The antibodies used, diluted in primary antibody diluents (P0023A, Beyotime), included: rabbit anti-phospho-ErbB4 (bs-3220R, Bioss; 1:1,000), rabbit anti-ErbB4 (AF6807, Beyotime; 1:1,000), rabbit anti-phospho-Akt1 (AF1546, Beyotime; 1:1,000), mouse anti-Akt1 (sc-377457, Santa Cruz Biotech; 1:1,000), rabbit anti-Nrf2 (AF7623, Beyotime; 1:1,000), rabbit anti-TFRC (AF8136, Beyotime; 1:1,000), rabbit anti-SLC7A11 (GB115276, Servicebio Biotech; 1:1,000), rabbit anti-GPX4 (AF7020, Beyotime; 1:1,000), rabbit anti-Lamin B1 (12987-1-AP, Proteintech, 1:5,000), rabbit anti-p53 (AF5258, Beyotime; 1:1,000), mouse anti-p21 (AP021, Beyotime; 1:1,000), rabbit anti-p16 (bs-23797R, Bioss Biotech, Beijing, China; 1:1,000), and rabbit anti-GAPDH (GB15002, Servicebio Biotech; 1:2000).

Techniques: Activation Assay, Inhibition, Expressing, Marker