anti cleaved poly  (Bioss)

Journal: Neural Regeneration Research

Article Title: Treadmill exercise in combination with acousto-optic and olfactory stimulation improves cognitive function in APP/PS1 mice through the brain-derived neurotrophic factor- and Cygb-associated signaling pathways

doi: 10.4103/NRR.NRR-D-23-01681

Figure Lengend Snippet: Multifactor stimulation increases Bcl-2, PARP, and LC3B expression and inhibits Bax and caspase-9 expression in the hippocampus of APP/PS1 mice. (A) Western blot analysis of Bcl-2, PARP, Bax, Caspase-9, and LC3B expression in the mouse hippocampus. β-Actin was used as a loading control. (B, C) Quantification of the band intensities showed that Bcl-2 (B, F (7, 16) = 67.55, P < 0.0001) and PARP (C, F (7, 16) = 71.51, P < 0.0001) were expressed at higher levels in the AD-stim group than in the AD-Ctrl group in both 4- and 6-month-old mice. n = 3 brains per group. (D, E) Quantitative analysis demonstrating that Bax (D, F (7, 16) = 33.40, P < 0.0001) and Caspase-9 (E, F (7, 16) = 118.3, P < 0.0001) expression levels were significantly lower in the AD-Stim group compared with the AD-Ctirl group in both 4- and 6-month-old mice. n = 3 brains per group. (F) LC3B was expressed at significantly higher levels in the AD-Stim group compared with the AD-Ctrl group in both 4- and 6-month-old mice.

Article Snippet: The following primary antibodies were used: postsynaptic density protein-95 (PSD-95, rabbit, 1:1000, Cell Signaling Technology, Cat# 3409, RRID: AB_1264242), synaptophysin (SYP, mouse, 1:1000, Abcam, Cat# ab8049, RRID: AB_2198854), doublecortin (DCX, rabbit, 1:1000, Cell Signaling Technology, Cat# 4604, RRID: AB_561007), NeuN (rabbit, 1:1000, Abcam, Cat# ab177487, RRID: AB_2532109), B cell lymphoma-2 (Bcl-2, rabbit, 1:1000, Abcam, Cat# ab182858, RRID: AB_2715467), Bcl2-associated X (Bax, rabbit,1:1000, Abcam, Cat# ab32503, RRID: AB_725631), Caspase-9 (rabbit, 1:1000, Abcam, Cat# ab185719), poly ADP-ribose polymerase (PARP, mouse, 1:1000, Cell Signaling Technology, Cat# 9541, RRID: AB_331426), brain-derived neurotrophic factor (BDNF, rabbit, 1:1000, Abcam, Cat# ab108319, RRID: AB_10862052), phosphorylated Ser473-Akt (p-Ser473-Akt, rabbit, 1:1000, Cell Signaling Technology, Cat# 4060, RRID: AB_2315049), phosphorylated Thr308-Akt (p-Thr308-Akt, rabbit, 1:500, Cell Signaling Technology, Cat# 13038, RRID: AB_2629447), microtubule-associated protein 1 light chain-3B (LC3B, rabbit, 1:2000, Novus Biologicals, Littleton, CO, USA, Cat# NB100-2220, RRID: AB_10003146), Cytoglobin (Cygb, mouse1:2000, Proteintech, Wuhan, Hubei Province, China, Cat# 60228-1-Ig, RRID: AB_11182383), glutathione peroxidase 4 (GPX4, rabbit, 1:1000, Abcam, Cat# ab125066, RRID: AB_10973901), solute carrier family 7 member 11 (SLC7A11, mouse, 1:1000, Cell Signaling Technology, Cat# 98051, RRID: AB_2800296), yes-associated protein 1 (YAP1, rabbit, 1:1000, Cell Signaling Technology, Cat# 14074, RRID: AB_2650491), and beta-actin (rabbit, 1:10,000, Abcam, Cat# ab8227, RRID: AB_2305186).

Techniques: Expressing, Western Blot, Control

Journal: Chinese Medical Journal

Article Title: LGR5 interacts with HSP90AB1 to mediate enzalutamide resistance by activating the WNT/β-catenin/AR axis in prostate cancer

doi: 10.1097/cm9.0000000000003538

Figure Lengend Snippet: Figure 3: LGR5 reduces sensitivity to ENZ. (A, B) Cell activity after treatment with ENZ was detected when LGR5 was overexpressed in 22Rv1 (A) and (B) C4-2 (n = 3). (C,D) Flow cytometry analysis of apoptosis in 22Rv1 (C) and C4-2 (D) cells treated with ENZ after LGR5 knockdown. (E, F) Flow cytometry analysis of apoptosis in 22Rv1 (E) and C4-2 (F) cells treated with ENZ after LGR5 overexpression. (G, H) Quantitative analysis of mean apoptosis rate (n = 3). (I, J) The Western blotting analysis was used to detect the changes of apoptosis indicator proteins in cells treated with ENZ after LGR5 knockdown or overexpression (n = 3). (K) Representative images of LGR5 overexpressing and control tumor mice treated with ENZ (10 mg/kg). (L) Tumor weight was measured after surgical resection of the tumor (n = 5). *P <0.05, †P <0.01, ‡P <0.001. ENZ: Enzalutamide; FITC: Fluorescein isothiocyanate; PARP: Poly (ADP-ribose) polymerase; PI: Propidium iodide; LGR5: Leucine-rich repeated G-protein-coupled receptor 5.

Article Snippet: Western blotting analysis was performed according to previously reported methods[32] with the following primary antibodies: LGR5 (#A10545, ABclonal), HSP90AB1 (#11405-1-AP, Proteintech), α-tubulin (#11224-1-AP, Proteintech), p-GSK3β (#67558-1-Ig, Proteintech), GSK-3β (#22104-1-AP, Proteintech), β-catenin (#51067- 2-AP, Proteintech), c-myc (#10828-1-AP, Proteintech), cyclin D1 (#26939-1-AP, Proteintech), GAPDH (#10494- 1-AP, Proteintech), cleaved poly (ADP-ribose) polymerase (PARP) (#5625T, CST, Boston, MA, USA), cleaved caspase 3 (#9664T, CST), and cleaved caspase 9(#7237T, CST).

Techniques: Activity Assay, Flow Cytometry, Knockdown, Over Expression, Western Blot, Control

Journal: Chinese Medical Journal

Article Title: LGR5 interacts with HSP90AB1 to mediate enzalutamide resistance by activating the WNT/β-catenin/AR axis in prostate cancer

doi: 10.1097/cm9.0000000000003538

Figure Lengend Snippet: Figure 5: HSP90AB1 knockdown reversed LGR5-induced ENZ resistance. (A) The major protein of WNT/β-catenin signaling pathway levels was detected by Western blotting analysis in LGR5-overexpressing cells combined with HSP90AB1 siRNA (n = 3). (B, C) Flow cytometry was used to detect the apoptosis of LGR5 overexpression cells treated with ENZ after HSP90AB1 knockdown (n = 3). (D) The Western blotting analysis was used to detect the changes of apoptosis indicator proteins in LGR5-overexpressing cells treated with ENZ after HSP90AB1 knockdown (n = 3). (E, F) Quantitative analysis of mean apoptosis rate in LGR5 overexpression cells treated with ENZ after HSP90AB1 knockdown (n = 3). *P <0.05, †P <0.01, ‡P <0.001. ENZ: Enzalutamide; GSK: Glycogen synthase kinase; HSP90AB1: Heat Shock Protein 90kDa Alpha B1; LGR5: Leucine-rich repeated G-protein-coupled receptor 5; PARP: Poly (ADP-ribose) polymerase; PI: Propidium iodide; siRNA: small interfering RNA; WNT: Wingless/integrated.

Article Snippet: Western blotting analysis was performed according to previously reported methods[32] with the following primary antibodies: LGR5 (#A10545, ABclonal), HSP90AB1 (#11405-1-AP, Proteintech), α-tubulin (#11224-1-AP, Proteintech), p-GSK3β (#67558-1-Ig, Proteintech), GSK-3β (#22104-1-AP, Proteintech), β-catenin (#51067- 2-AP, Proteintech), c-myc (#10828-1-AP, Proteintech), cyclin D1 (#26939-1-AP, Proteintech), GAPDH (#10494- 1-AP, Proteintech), cleaved poly (ADP-ribose) polymerase (PARP) (#5625T, CST, Boston, MA, USA), cleaved caspase 3 (#9664T, CST), and cleaved caspase 9(#7237T, CST).

Techniques: Knockdown, Western Blot, Flow Cytometry, Over Expression, Small Interfering RNA