bh3 interacting domain death agonist 10988 1 ap  (Proteintech)

Journal: Translational Cancer Research

Article Title: Neochlorogenic acid inhibits gastric cancer cell growth through apoptosis and cell cycle arrest

doi: 10.21037/tcr-2025-696

Figure Lengend Snippet: Western blot analysis reveals NCA-mediated modulation of apoptosis-associated proteins in HGC-27 and NUGC-3 cells. Following a 48-hour treatment with control or NCA, the expression of BAX, CASP3, BID, CYCS, and BCL2 was examined by Western blotting. β-actin was used as a loading control. Protein levels were normalized to β-actin and expressed as fold changes relative to the untreated control group. Bar graphs depict the quantification of relative protein levels. Data represent mean ± standard deviation from three independent experiments (n=3). Statistical significance was determined by Student’s t -test. *, P<0.05; **, P<0.01; ***, P<0.001. BAX, BCL2-associated X protein; BCL2, B-cell lymphoma 2; BID, BH3 interacting domain death agonist; CASP3, cysteinyl aspartate specific proteinase 3; CYCS, cytochrome c, somatic; NCA, neochlorogenic acid.

Article Snippet: The membranes were then incubated overnight at 4 °C with primary antibodies for BCL2 (B-cell lymphoma 2) (Cat. No. bs-0032R; Bioss, Beijing, China), BID (BH3 interacting domain death agonist) (Cat. No. 10988-1-AP), CYCS (cytochrome c, somatic) (Cat. No. 10993-1-AP) from Proteintech (Wuhan, China), as well as β-actin (Cat. No. 81115-1-RR, Proteintech, Wuhan, China) and BAX (BCL2-associated X protein) (Cat. No. 50599-2-Ig, Proteintech, Wuhan, China).

Techniques: Western Blot, Control, Expressing, Standard Deviation

Journal: Folia histochemica et cytobiologica

Article Title: Electroacupuncture stimulation inhibited astrogliosis and microglia polarisation to alleviate spinal cord injury via Janus kinase 2/signal transducer and activator of transcription 3 signalling pathway.

doi: 10.5603/fhc.104273

Figure Lengend Snippet: Figure 2. Electroacupuncture (EA) protected neurons against spinal cord injury (SCI)-induced apoptosis. Tissues were obtained and processed as described for Fig. 1 and in Methods. A. Nissl staining of spinal cord. Upper panel shows full cross sections of spinal cord, scale bar = 500 μm. Lower panel shows partial enlargement of sections, scale bar = 50 μm; B. Representative images showing immunofluo- rescence of TUNEL+/NeuN+ in rat spinal cord, scale bar = 50 μm; C. Quantification of percentage of TUNEL and NeuN dual-positive cells in spinal cord; D. Left panel shows immune blots of B-cell lymphoma-2 associated X protein (Bax), BH3-interacting domain death agonist (Bid), and B-cell lymphoma-2 (Bcl-2). Right panel shows quantification of Bax, Bid, and Bcl-2 protein expression in lysates of spinal cord. Numeric data refers to Figs. 2C, D, results expressed as mean ± standard deviation, n = 6 in each group. *P < 0.05, **P < 0.01, ***P < 0.001, API — 4’,6-diamidino-2-phenylindole; Ns — not significant.

Article Snippet: The PVDF membrane was blocked with a membrane sealing solution (Solarbio) at RT for 1 h. Primary antibodies [B-cell lymphoma-2 (Bcl-2; 1:1,000, Proteintech), BH3-interacting domain death agonist (Bid; 1:1,500, Proteintech), B-cell lymphoma-2 associated X protein (Bax; 1:5,000, Proteintech), JAK2 (1:1,500, Affinity), STAT3 (1:2,000, Proteintech), Phospho-JAK2 (p-JAK2; 1:800, ABclonal, Wuhan, China), p-STAT3 (1:1,000, Affinity), and glyceraldehyde phosphate dehydrogenase (GAPDH; 1:10,000, Proteintech)] were incubated with the PVDF membranes overnight at 4°C and washed with Tris-buffered saline with tween-20 (TBS-T) buffer.

Techniques: Staining, TUNEL Assay, Expressing, Standard Deviation

Journal: Journal of Translational Medicine

Article Title: Doxorubicin synergizes bortezomib-induced multiple myeloma cell death by inhibiting aggresome formation and augmenting endoplasmic reticulum/Golgi stress and apoptosis

doi: 10.1186/s12967-024-05920-2

Figure Lengend Snippet: Cotreatment with doxorubicin increases misfolded protein accumulation, enhances ER/Golgi stress responses and promotes apoptosis/necroptosis in bortezomib-treated cells. A Cotreatment with doxorubicin increases the level of ubiquitinated proteins in bortezomib-treated cells. U266B1 cells exposed to bortezomib or the combination of bortezomib and doxorubicin were analyzed for misfolded protein levels by Western blot using anti-ubiquitin antibodies. The relative protein levels were quantified and plotted with statistical significance. B Cotreatment with doxorubicin elevates the protein levels of factors that promote ER stress-dependent apoptosis in bortezomib-treated cells. U266B1 cells exposed to bortezomib or the combination were analyzed by Western blot for the levels of ER stress response regulators, including IRE1, pIRE-S724, GRP78, CHOP, ATF3, and ATF4. The relative protein levels were plotted with statistical significance. C Cotreatment with doxorubicin increases the protein levels of factors involved in Golgi stress-dependent apoptosis in bortezomib-treated cells. Western blot analysis was performed on U266B1 cells treated with bortezomib or the combination, focusing on Golgi stress response regulators such as ARF4, CREB3, ETS1, ETS2, and HSP47. The relative protein levels were quantified and plotted. D Cotreatment with doxorubicin enhances the levels of proapoptotic factors in Bortezomib-treated cells. U266B1 cells were analyzed by Western blot for proapoptotic factors such as Bad, pBad (S112), Bax, Bik, Bim, BID, Bak, and Puma. The relative protein levels were plotted with statistical significance. E Cotreatment with doxorubicin increases the levels of active caspases in bortezomib-treated cells. Western blot analysis was performed on U266B1 cells exposed to Bortezomib or the combination, measuring the levels of the proform and cleaved active forms of caspases 3, 6, 7, 9, and PARP. The relative protein levels were plotted with statistical significance. F Cotreatment with doxorubicin elevates the protein level of RIPK1, a key factor promoting necroptosis, in Bortezomib-treated cells. U266B1 cells were analyzed by Western blot for necroptosis regulators, including RIPK1, RIPK2, and RIPK3. The relative protein levels were quantified and plotted with statistical significance. Statistical significance was determined using t-tests, with *p < 0.05, **p < 0.01, and ***p < 0.001. All experiments were performed independently three times

Article Snippet: Antibodies against p-38 (#8690P), phosphorylated-p38 (#4511P), Bcl-2-associated death promoter (Bad) (#9239), phosphorylated Bad serine 112 (#5284), Bcl-2-associated X protein (Bax) (#5023), Bcl-2-interacting killer (Bik) (#4592), Bcl-2-interacting mediator of cell death (Bim) (#2933), BH3-interacting domain death agonist (BID) (#2002), Bcl-2 homologous antagonist/killer (Bak) (#12105), p53 upregulated modulator of apoptosis (Puma) (#12450), Caspase 3 (#14220), cleaved Caspase 3 (#9664), Caspase 7 (#12827), cleaved Caspase 7 (#8438), Caspase 9 (#9508), cleaved Caspase 9 (#52873), poly(ADP-ribose) polymerase (PARP) (#9542), cleaved PARP (#5625), receptor-interacting serine/threonine-protein kinase 1 (RIPK1) (#3493), mixed lineage kinase domain-like protein (#14993), and RIPK3 (#13526) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Western Blot, Ubiquitin Proteomics

Journal: Journal of Translational Medicine

Article Title: Characterization of the proteome of stable and unstable carotid atherosclerotic plaques using data-independent acquisition mass spectrometry

doi: 10.1186/s12967-023-04723-1

Figure Lengend Snippet: Validation of ferroptosis- and lipid metabolism-associated DEPs using IHC in a validation cohort. A Representative images (see Additional file : Figure S4a,b for 500 µm images) of immunohistochemical staining of TFR1, TF, AIFM2, DPP4, and GCLC proteins in stable and unstable plaques in the plaque fibrous cap region (black arrows) and immunohistochemical staining for SLC1A5, BID, and APOA5 proteins in the plaque lipid core region (black arrows). B In unstable plaques, the levels of TFR1, TF, AIFM2, DPP4, GCLC, SLC1A5, BID, and APOA5 were significantly increased, while other differences were not statistically significant. IHC immunohistochemistry. TFR1 transferrin receptor protein 1; TF transferrin; AIFM2 apoptosis-inducing factor 2; DPP4 dipeptidyl peptidase 4; GCLC glutamate-cysteine ligase catalytic. SLC1A5 solute carrier family 1, member 5; BID BH3 interacting-domain death agonist; APOA5, apolipoprotein A-V; CETP cholesteryl ester transfer protein. *P < 0.05; **P < 0.01 Student’s t test (two-tailed distribution)

Article Snippet: Primary antibodies for IHC against solute carrier family 1, member 5 (SLC1A5) (rabbit polyclonal, 20350-1-AP), apoptosis-inducing factor 2 (AIFM2) (rabbit polyclonal, 20886–1-AP), BH3 interacting-domain death agonist (BID) (rabbit polyclonal, 10988-1-AP), dipeptidyl peptidase 4 (DPP4) (rabbit polyclonal, 29403-1-AP), transferrin receptor protein 1 (TFR1) (mouse monoclonal, 66180–1-Ig), transferrin (TF) (rabbit polyclonal, 17435-1-AP), glutamate–cysteine ligase catalytic (GCLC) (rabbit polyclonal, 12601–1-AP), cholesteryl ester transfer protein (CETP) (rabbit polyclonal, 13459-1-AP), and apolipoprotein A-V (APOA5) (rabbit polyclonal, 18019-1-AP) were purchased from Proteintech (China).

Techniques: Biomarker Discovery, Immunohistochemical staining, Staining, Immunohistochemistry, Two Tailed Test

Journal: Journal of Translational Medicine

Article Title: Characterization of the proteome of stable and unstable carotid atherosclerotic plaques using data-independent acquisition mass spectrometry

doi: 10.1186/s12967-023-04723-1

Figure Lengend Snippet: The differently expressed proteins (DEPs) that were validated using immunohistochemistry (all up regulated) (unstable/stable)

Article Snippet: Primary antibodies for IHC against solute carrier family 1, member 5 (SLC1A5) (rabbit polyclonal, 20350-1-AP), apoptosis-inducing factor 2 (AIFM2) (rabbit polyclonal, 20886–1-AP), BH3 interacting-domain death agonist (BID) (rabbit polyclonal, 10988-1-AP), dipeptidyl peptidase 4 (DPP4) (rabbit polyclonal, 29403-1-AP), transferrin receptor protein 1 (TFR1) (mouse monoclonal, 66180–1-Ig), transferrin (TF) (rabbit polyclonal, 17435-1-AP), glutamate–cysteine ligase catalytic (GCLC) (rabbit polyclonal, 12601–1-AP), cholesteryl ester transfer protein (CETP) (rabbit polyclonal, 13459-1-AP), and apolipoprotein A-V (APOA5) (rabbit polyclonal, 18019-1-AP) were purchased from Proteintech (China).

Techniques: Immunohistochemistry, Biomarker Discovery, Significance Assay

Journal: Journal of Translational Medicine

Article Title: Characterization of the proteome of stable and unstable carotid atherosclerotic plaques using data-independent acquisition mass spectrometry

doi: 10.1186/s12967-023-04723-1

Figure Lengend Snippet: Hypothetical characterization of altered molecular mechanisms in cells in the fibrous cap and lipid core regions of unstable carotid plaques. The expression of key proteins of ferroptosis and lipid metabolism is significantly increased in patients with unstable plaques, and there are mechanisms associated with both the promotion and inhibition of iron death. This possibly indicates that cells in different regions of the plaque are regulated by key proteins of ferroptosis that subsequently lead to increased plaque instability and expansion of the necrotic core. TFR1 transferrin receptor protein 1; TF transferrin; AIFM2 apoptosis-inducing factor 2; DPP4 dipeptidyl peptidase 4; GCLC glutamate-cysteine ligase catalytic; SLC1A5 solute carrier family 1, member 5; BID BH3 interacting-domain death agonist; APOA5 apolipoprotein A-V; CETP cholesteryl ester transfer protein; GPX4 glutathione peroxidase 4; GSH glutathione; CoQ10 coenzyme Q10; ROS reactive oxygen species; HDL high-density lipoprotein; ox-LDL oxidized low-density lipoprotein

Article Snippet: Primary antibodies for IHC against solute carrier family 1, member 5 (SLC1A5) (rabbit polyclonal, 20350-1-AP), apoptosis-inducing factor 2 (AIFM2) (rabbit polyclonal, 20886–1-AP), BH3 interacting-domain death agonist (BID) (rabbit polyclonal, 10988-1-AP), dipeptidyl peptidase 4 (DPP4) (rabbit polyclonal, 29403-1-AP), transferrin receptor protein 1 (TFR1) (mouse monoclonal, 66180–1-Ig), transferrin (TF) (rabbit polyclonal, 17435-1-AP), glutamate–cysteine ligase catalytic (GCLC) (rabbit polyclonal, 12601–1-AP), cholesteryl ester transfer protein (CETP) (rabbit polyclonal, 13459-1-AP), and apolipoprotein A-V (APOA5) (rabbit polyclonal, 18019-1-AP) were purchased from Proteintech (China).

Techniques: Expressing, Inhibition