Journal: Life sciences in space research

Article Title: Comparison of signaling profiles in the low dose range following low and high LET radiation.

doi: 10.1016/j.lssr.2020.02.002

Figure Lengend Snippet: Fig. 1. Average total fold intensity of phospho-protein signal as compared to median control value. Colored bars (dark blue = 0.5 Gy, light blue = 0.1 Gy and green = 0.05 Gy) indicate values significantly different from controls (yellow). Significance bars are shown for all significant differences between doses and 0 (p ≤0.05). Average fold intensity levels are shown at 2 h post radiation for γH2AX (A), pATF2 (B) and pSMC1 (C). Persistent effects are shown at 24 h for γH2AX (D), pATF2 (E) and pSMC1 (F). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: For staining, 0.5 × 106 fixed cells were washed in phosphate buffered saline (PBS), resuspended in blocking buffer (2% FBS/PBS) and incubated for 1 h with primary antibody on ice, with resuspension of the pellet every 15 min. Primary antibodies used in flow cytometry include mouse monoclonal γH2AXSer139 (1:800 dilution) and pSMC1Ser957 (1:800 dilution) from Millipore (Temecula, CA) and rabbit polyclonal pATF2 Ser490/498 (1:1000 dilution) from Rockland, Inc. (Gilbertsville, PA).

Techniques: Control

Journal: Life sciences in space research

Article Title: Comparison of signaling profiles in the low dose range following low and high LET radiation.

doi: 10.1016/j.lssr.2020.02.002

Figure Lengend Snippet: Fig. 3. Average total fold pATF2 intensity over median control level versus fluence for various radiation qualities. Average fold intensity levels are shown at 2 h post radiation for Si ions (A),Fe ions (C) and Ti ions (E). Persistent effects are shown at 24 h for Si ions (B), Fe ions (D) and Ti ions (F).

Article Snippet: For staining, 0.5 × 106 fixed cells were washed in phosphate buffered saline (PBS), resuspended in blocking buffer (2% FBS/PBS) and incubated for 1 h with primary antibody on ice, with resuspension of the pellet every 15 min. Primary antibodies used in flow cytometry include mouse monoclonal γH2AXSer139 (1:800 dilution) and pSMC1Ser957 (1:800 dilution) from Millipore (Temecula, CA) and rabbit polyclonal pATF2 Ser490/498 (1:1000 dilution) from Rockland, Inc. (Gilbertsville, PA).

Techniques: Control

Journal: Life sciences in space research

Article Title: Comparison of signaling profiles in the low dose range following low and high LET radiation.

doi: 10.1016/j.lssr.2020.02.002

Figure Lengend Snippet: Fig. 6. Average total fold intensity of pATF2 signal intensity divided by fluence and graphed versus LET. Average fold intensity levels are shown at 2 h post radiation for 0.05 Gy (A), 0.1 Gy (B) and 0.5 Gy (C). Persistent effects are shown at 24 h for 0.05 Gy (D), 0.1 Gy (E) and 0.5 Gy (F).

Article Snippet: For staining, 0.5 × 106 fixed cells were washed in phosphate buffered saline (PBS), resuspended in blocking buffer (2% FBS/PBS) and incubated for 1 h with primary antibody on ice, with resuspension of the pellet every 15 min. Primary antibodies used in flow cytometry include mouse monoclonal γH2AXSer139 (1:800 dilution) and pSMC1Ser957 (1:800 dilution) from Millipore (Temecula, CA) and rabbit polyclonal pATF2 Ser490/498 (1:1000 dilution) from Rockland, Inc. (Gilbertsville, PA).

Techniques:

Journal: eLife

Article Title: Dependency of human and murine LKB1-inactivated lung cancer on aberrant CRTC-CREB activation

doi: 10.7554/eLife.66095

Figure Lengend Snippet:

Article Snippet: The following antibodies were used for western blotting: anti-CRTC1 (Cat #600-401-936, Rabbit) from Rockland Immunochemicals Inc; anti-CRTC2 (Cat #A300-637A, Rabbit) and anti-PDE4D (Cat #A302-744A, Rabbit) from Bethyl Laboratories; anti-CRTC3 (Cat #2720, Rabbit), anti-LKB1 (Cat #3050, Rabbit) from Cell Signaling Technology; anti-HDCA1 (Cat # sc7872, rabbit) from Santa Cruz Biotechnology, anti-β-TUBULIN (Cat #1878, Rabbit) from Epitomics; and anti-β-ACTIN (Cat #A5316, Mouse) from Sigma-Aldrich.

Techniques: Recombinant, Plasmid Preparation, Sequencing, Clone Assay, Control, Cell Culture, Transfection, RNA Extraction, Reverse Transcription, SYBR Green Assay, Extraction, Reporter Assay, Software

Journal: Neurochemical Research

Article Title: Morin Improves Cognitive Deficits in an in Vivo Model of Vascular Dementia by Modulating the N-methyl-D-aspartate Receptor Signaling Pathways

doi: 10.1007/s11064-026-04717-7

Figure Lengend Snippet: Morin modulated the expression of NMDA receptors in the hippocampus of VaD rats. a the expression levels of NR1 ; b the expression levels of NR2A ; c the expression levels of NR2B ; d the expression levels of NR1 protein; e the expression levels of NR2A protein; f the expression levels of NR2B protein; g protein levels of p-CREB; h protein levels of p-CAMK2A; i protein levels of p-CAMK2D. Protein levels of NR1, NR2A, and NR2B were quantified by ELISA. Data are presented as mean ± SD ( n = 8 per group). Statistical analysis was performed by one-way ANOVA with Tukey’s post-hoc test (data met assumptions of normality and homoscedasticity)/Kruskal-Wallis with Dunn’s test (data did not meet assumptions). * indicates a significant difference from the Sham group; # indicates a significant difference 2VO group; *# indicates a significant difference from both the Sham and 2VO groups, with a p-value of less than 0.05 considered statistically significant

Article Snippet: The phosphorylation status of another key NMDAR downstream effector, CREB at Ser133 (p-CREB), was quantified in hippocampal homogenates using a commercial ELISA kit (Phospho-CREB [Ser133] ELISA Kit, Boster Bio, #EKC2382).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Peripheral amylin modulation rebalances brain glycolysis and Tau-Ser214 phosphorylation via cAMP-PKA signaling

doi: 10.1016/j.isci.2026.115157

Figure Lengend Snippet: ATF4 signaling, glucose transporters and amino acid responses in low vs. high brain amylin under metabolic stress (A and B) Top seven canonical pathways from ingenuity pathways analysis (A) and (B) top five GO biological processes enriched in hA i−OFF vs. hA ON brains. (C) Brain ATF4 levels in hA ON , hA i−OFF , hA OFF , and hA i−ON mice. Main effect of animal model, p = 0.0049. (D) Aspartate to asparagine ratio in brain tissues from hA ON , hA i−OFF , hA OFF , and hA i−ON mice. Main effect of animal model, p < 0.0001. (E) Correlation between brain amylin and ATF4, with mice stratified by normal (126.1 ± 3.15 mg/dL) or high (154.4 ± 5.39 mg/dL) glucose. (F–I) Statistical modeling comparing molecular outcomes (cAMP and ATF4) of changes in plasma amylin vs. insulin levels in same mice. (J and K) GLUT1 and GLUT3 levels in brain tissues from hA ON , hA i−OFF , hA OFF , and hA i−ON mice as a function of blood glucose levels. Measurements were performed in duplicate. Results are shown as individual data points and/or mean ± s.e.m. and statistical analyses were performed using One-way ANOVA followed by Tukey’s multiple-comparisons test (C and D).

Article Snippet: ATF4 , Proteintech , KE00147.

Techniques: Animal Model, Clinical Proteomics

Journal: iScience

Article Title: Peripheral amylin modulation rebalances brain glycolysis and Tau-Ser214 phosphorylation via cAMP-PKA signaling

doi: 10.1016/j.isci.2026.115157

Figure Lengend Snippet: Pancreatic amylin regulation reduces amylin-cAMP-PKA overactivation and tau-Aβ coupling: molecular evidence (A and B) Representative confocal microscopy images comparing neuronal pS214-tau in cortical sections from hA ON vs. hA OFF mice ( n = 5 slices/mouse from n = 3 mice/group). (C–F) In the same mice as in , , , , , , and , average brain amylin level vs. average pS214-tau (C) and average brain tissue PKA activity vs. average pS214-tau (D) along with correlation analyses between brain tissue Aβ 42 and pT31-tau in hA ON vs. hA i−OFF mice (E) and hA OFF vs. hA i−ON mice (F). (G) Heat maps comparing the brain tissue levels of inflammatory cytokines (IL-1β, IL-6, TNF-α, IFN-γ, and CXCL1) in hA i−OFF mice vs. hA ON littermates. (H) Same as in (G) for hA i−ON mice compared to hA OFF littermates. (I) In same mice, principal component (PC) analysis of amylin, AD markers (Aβ 40, Aβ 42 , pTau, and total tau), neuroinflammation (IL-1β, IL-6, TNF-α, IFN-γ, and CXCL1), and metabolic covariates (glucose, G6P, cAMP, and ATF4). (J) PC loading plot suggesting that clustering is largely driven by contributions of amylin, cAMP and Aβ covariates to the main principal component (PC1), whereas G6P and tau pathology drive the variation on PC2. (K) Schematic diagram (using BioRender): toggling amylin secretion under pre-diabetic stress bidirectionally regulates brain glucose metabolism and memory. Excess amylin amplifies amylin-cAMP-PKA signaling, suppresses astrocytic glycolysis and promotes tau phosphorylation, tau-Aβ coupling and neuroinflammation, whereas restricting brain amylin preserves metabolic function and memory.

Article Snippet: ATF4 , Proteintech , KE00147.

Techniques: Confocal Microscopy, Activity Assay, Phospho-proteomics

Journal: International Journal of Molecular Sciences

Article Title: Strontium Attenuates Hippocampal Damage via Suppressing Neuroinflammation in High-Fat Diet-Induced NAFLD Mice

doi: 10.3390/ijms241210248

Figure Lengend Snippet: Sr restrained the HFD-induced apoptosis by altering expression levels of proteins related to the ERS pathway. ( A ) Western blot analysis of caspase-3, GRP78, IRE1α, p-IRE1α, XBP1, eIF2α, p-eIF2α, ATF4, ATF6, CHOP, and β-actin. ( B – K ) Relative protein expression of caspase-3 ( B ), GRP78 ( C ), IRE1α ( D ), p-IRE1α ( E ), XBP1 ( F ), eIF2α ( G ), p-eIF2α ( H ), ATF4 ( I ), ATF6 ( J ), and CHOP ( K ) in the hippocampi of each group of mice was examined through Western blotting ( n = 6 per group). Data were normalized with respect to the band of β-actin: the expression of target protein = the intensity of target protein band/the intensity of β-actin band. Results are shown as the ratio of the experimental group to the control group, and the values of the control group were taken as 1. All data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Subsequently, the following primary antibodies were used to incubate the membranes overnight at 4 °C: rabbit anti- NF-κB (#8242), rabbit anti- p38 (#9212), rabbit anti- ERK (#9102), rabbit anti-phospho- ERK ( p-ERK , #4370), rabbit anti-phospho- p38 ( p-p38 , #4511), and anti- caspase-3 (#9662) (purchased from Cell Signaling Technology (Danvers, MA, USA)); mouse anti- ATF6 (EM1701-94) (purchased from Hangzhou Huaan Biotechnology Co., Ltd., Hangzhou, China); rabbit anti- XBP1 (A1731), rabbit anti-phospho- NF-κB ( p- NF-κB , AP0475), rabbit anti- GRP78 (A0241), and mouse anti- β-actin (purchased from Wuhan ABclonal Technology Co., Ltd., Wuhan, China); rabbit anti- eIF2α (ab115822), rabbit anti- TLR4 (ab13556), and rabbit anti-phospho- eIF2α ( p-eIF2α , ab32157) (purchased from Abcam (Cambridge, MA, USA)); and rabbit anti- CHOP (BM4962), anti-phospho- IRE1α ( p-IRE1α , BM4444), rabbit anti- ATF4 (BM5179), and rabbit anti- IRE1α (A00683-1) (purchased from Wuhan BOSTER Biological Technology Co., Ltd., Wuhan, China).

Techniques: Expressing, Western Blot, Control

Journal: FEBS Open Bio

Article Title: Pyruvate alleviates high glucose‐induced endoplasmic reticulum stress and apoptosis in HK‐2 cells

doi: 10.1002/2211-5463.12834

Figure Lengend Snippet: Pyr effects on ER stress‐related protein and ROS level. HK‐2 cells were treated with Pyr (0.5 m m ) and/or HG (30 m m glucose) for 3 days. The protein expressions (A) of GRP78, CHOP, ATF4 and p‐EIF2α of HK‐2 cell were detected by western blot analyses. The percentages of GRP78 (B), CHOP (C), ATF4 (D) and p‐EIF2α (E)/β‐actin in the bar graphs were quantified by imagej software. Expressions of GRP78 (B), CHOP (C), ATF4 (D) and p‐EIF2α (E) were decreased after Pyr treatments, which ascertained that exogenous Pyr ameliorated the ER stress in HK‐2 cells. (F) HK‐2 cells were treated with Pyr (0.5 m m ) and/or HG (30 m m glucose) for 3 days. Further, HG‐induced ROS increases were inhibited by Pyr treatments in HK‐2 cells. Con: cells were treated with 5 m m glucose in the DMEM; HG: cells were treated with 30 m m glucose in the DMEM; HG + Pyr: cells were treated with 30 m m glucose and 0.5 m m Pyr in the DMEM; Pyr: cells were treated with 5 m m glucose and 0.5 m m Pyr in the DMEM. Values were means ± SEM ( n = 5). Data were analyzed by one‐way ANOVA. * P < 0.05; ** P < 0.01.

Article Snippet: The primary antibodies against Bcl‐2, BAX, Caspase‐3, glucose‐regulated protein 78 (GRP78), C/EBP homologous protein (CHOP), activating transcription factor 4 (ATF4) and phosphorylate α‐subunit of eukaryotic initiation factor 2 (p‐EIF2α) and the secondary antibodies (anti‐mouse IgG, anti‐rabbit IgG) were obtained from Proteintech Group, Inc (Chicago, IL, USA), and the antibody against p‐EIF2α was purchased from CST (Danvers, MA, USA).

Techniques: Western Blot, Software