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: Antioxidants

Article Title: Dimercaprol Reprograms Intestinal Redox Homeostasis and Organelle Crosstalk to Combat Iron-Induced Gut Dysbiosis Through NRF2/HO-1 Signaling

doi: 10.3390/antiox15030356

Figure Lengend Snippet: DP-dependent defense against FC-induced ER stress in IPEC-J2 cells is mediated by NRF2/HO-1 signaling. ( A , B ) Western blot band intensities and quantitative analyses indicate that FC exposure induces mitochondrial dysfunction and activates ER stress. This is evidenced by the notable increase in both protein and mRNA levels of canonical ER stress markers (ATF4, ATF6, CHOP, GRP78 and XBP1). DP attenuated this response, reducing the expression of these markers. In contrast, NRF2 knockdown abolished the protective effect of DP and further increased ER stress marker expression, indicating that DP-mediated ER stress suppression requires NRF2. ( C ) Fluorescence intensity confocal microscopy confirmed that NRF2 downregulation disrupts the colocalization of ER stress-related proteins, consistent with enhanced ER stress. Data are presented as mean ± SD for histogram ( A ) three replicates and ( B ) six replicates. Each group’s significant differences were compared with FC. ns, non-significant, * p < 0.01, ** p < 0.001, *** p < 0.0001, **** p < 0.00001.

Article Snippet: XBP1 , BOSTER (Wuhan, China) , PB9463 , WB.

Techniques: Western Blot, Expressing, Knockdown, Marker, Fluorescence, Confocal Microscopy

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: 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: Oncology reports

Article Title: TSPYL5 activates endoplasmic reticulum stress to inhibit cell proliferation, migration and invasion in colorectal cancer.

doi: 10.3892/or.2020.7639

Figure Lengend Snippet: Figure 6. Effects of TSPYL5 overexpression on ERS and its associated proteins. HCT116 and HT29 cells were transfected with pcDNA3.1‑TSPYL5 or empty pcDNA3.1, and subsequently divided into TSPYL5 and negative control (NC) groups, respectively. (A) Western blot analysis of the protein levels of caspase‑1, caspase‑3, bcl‑2‑like protein 4 (Bax), activating transcription factor 4 (ATF4) and CCAAT‑enhancer‑binding protein homologous protein (CHOP) in the HT29 and HCT116 cells in the TSPYL5 overexpression and NC groups. (B) Quantification of the levels of caspase‑1, caspase‑3, Bax, ATF4, and CHOP protein expression as determined by western blot analysis. *P<0.05, **P<0.01 and ***P<0.001 vs. the NC group. CRC, colorectal cancer; TSPYL5, testis‑specific protein Y‑encoded‑like 5.

Article Snippet: The membranes were then blocked with 5% skim milk and subsequently incubated overnight with primary antibodies against TSPYL5 (cat. no. ab203657, dilution: 1:800; Abcam), caspase‐1 (cat. no. ab62698, dilution: 1:800; Abcam), caspase‐3 (cat. no. ab49822, dilution: 1:500; Abcam), Bax (cat. no. M00183‐2, dilution: 1:1,000, Boster), ATF4 (cat. no. A00371, dilution: 1:500; Boster), CHOP (cat. no. A00311, dilution: 1:500; Boster), and GAPDH (cat. no. ab9485, dilution: 1:800; Abcam) at 4 ̊C.

Techniques: Over Expression, Transfection, Negative Control, Western Blot, Expressing