Journal: Pharmaceuticals

Article Title: Mazdutide Ameliorates Metabolic Dysfunction-Associated Steatotic Liver Disease by Modulating Endoplasmic Reticulum Stress, Improving Lipid Metabolism and Alleviating Inflammation

doi: 10.3390/ph19030371

Figure Lengend Snippet: Mazdutide suppresses ER stress via the PERK pathway in MASLD mouse liver tissue. ( A ) Representative immunohistochemical staining of GRP78 and CHOP in liver sections (scale bar = 50 μm, 200× magnification). ( B ) Representative Western blot images of key proteins in the PERK pathway from each experimental group. ( C ) Quantitative densitometric analysis of PERK and eIF2α phosphorylation (presented as the p-PERK/PERK and p-eIF2α/eIF2α ratios) and the protein levels of GRP78, ATF4, and CHOP. All values were normalized to β-actin. Data are presented as the mean ± SD ( n = 5). a p < 0.05 vs. the NCD group; b p < 0.05 vs. the HFD group. Animal groups are defined in .

Article Snippet: Primary antibodies for GRP78, ATF4, CHOP, and β-actin were purchased from Proteintech Group, Inc (Wuhan, China).

Techniques: Immunohistochemical staining, Staining, Western Blot, Phospho-proteomics

Journal: Pharmaceuticals

Article Title: Mazdutide Ameliorates Metabolic Dysfunction-Associated Steatotic Liver Disease by Modulating Endoplasmic Reticulum Stress, Improving Lipid Metabolism and Alleviating Inflammation

doi: 10.3390/ph19030371

Figure Lengend Snippet: Mazdutide inhibits the PERK-eIF2α-ATF4-CHOP pathway in MASLD cells. ( A ) Representative Western blot images showing the protein levels of GRP78, p-PERK, PERK, p-eIF2α, eIF2α, ATF4, CHOP, and β-actin. ( B ) Quantitative densitometric analysis of the p-PERK/PERK and p-eIF2α/eIF2α ratios and the protein levels of GRP78, ATF4, and CHOP. The protein band intensities were normalized to β-actin. Data are presented as the mean ± SD ( n = 3). a p < 0.05 vs. the NCD group; b p < 0.05 vs. the HFD group. Cell groups are as defined in .

Article Snippet: Primary antibodies for GRP78, ATF4, CHOP, and β-actin were purchased from Proteintech Group, Inc (Wuhan, China).

Techniques: Western Blot

Journal: The FASEB Journal

Article Title: Deficiency of IKKβ in neurons ameliorates Alzheimer's disease pathology in APP ‐ and tau‐transgenic mice

doi: 10.1096/fj.202201512r

Figure Lengend Snippet: FIGURE 10 Deficiency of IKKβ in neurons does not affect cognitive function and even increases apoptosis in the brains of tau-transgenic mice. In the water maze test, 9-month-old TautgIKKβfl/flCre+/− (IKKβ ko) and TautgIKKβfl/flCre−/− (IKKβ wt) littermate mice did not differ in traveling latency and distance to reach the escape platform during the training phase (A and B; two-way ANOVA, p > .05, n ≥ 6 per group), nor in the frequency with which the mice visited the region where the platform was previously located during the probe trial (C; t test, p > .05, n ≥ 6 per group). Western blotting was used to detect cleaved caspase-3, phosphorylation level of CREB, and the amount of synaptic structure proteins, Munc18-1, SNAP25, synaptophysin, and PSD-95 in the brain homogenate of tau-transgenic mice (D–M). Deficiency of IKKβ in neurons does not alter the protein levels of various synaptic proteins (E–H; t test, p > .05, n ≥ 4 per group), nor the ratio of phosphorylated (p-) to total (t-) CREB (J and K; t test, p > .05, n ≥ 6 per group). Surprisingly, IKKβ deficiency significantly increases the protein level of cleaved caspase-3 in the brains of 9-month-old tau-transgenic mice (M; t test, n ≥ 9 per group).

Article Snippet: Proteins were then transferred onto polyvinylidene difluoride (PVDF) or nitrocellulose membranes and incubated overnight at 4°C with the following antibodies: mouse monoclonal antibody against Aβ (clone W0- 2; Sigma- Aldrich); rabbit monoclonal antibodies against beclin1 (clone D40C5), phosphor- glycogen synthase kinase (GSK)- 3β (clone D3A4), GSK- 3β (clone 27C10), phospho- CREB (Ser133) (clone 87G3), CREB (clone 48H2), cleaved Caspase- 3 (clone 5A1E), PSD- 95 (clone D27E11), synaptophysin (clone D8F6H), SNAP25 (clone D110), and Munc18- 1 (clone D4O6V), and rabbit polyclonal antibodies against LC3B (Cat.- No: NB100- 2220; Novus Biologicals, Centennial, USA), SQSTM1/p62 (Cat.- No: 5114), phospho- p38- MAPK (Thr180/Tyr182) (Cat.- No: 9211), p38- MAPK (Cat.- No: 9212), and protein phosphatase type 2A (PP2A) catalytic subunit (Cat.- No: 2038) (all antibodies except anti- LC3B were bought from Cell Signaling Technology, Danvers, USA).

Techniques: Transgenic Assay, Western Blot, Phospho-proteomics

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: