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: Toxics

Article Title: Effects of Short-Term Exposure to Polystyrene Nanoplastics on the Nervous System: Calcium Homeostasis, BDNF and Synaptic Plasticity

doi: 10.3390/toxics14020178

Figure Lengend Snippet: Role of BDNF in the mechanism of synaptic plasticity impairment. All indicators were detected in neurons from the control and PS-NPs intervention groups. ( A ) Western blot analysis of CREB, BDNF, Cav3.1, and CaMKIIα in neurons, with GAPDH as the loading control. ( B ) Protein expression levels of CREB in the neurons. n = 3. ( C ) Protein expression levels of BDNF in the neurons. n = 3. ( D ) Calcium ion content in the neurons. n = 6. ( E ) Protein expression levels of the calcium channel Cav3.1 in neurons. ( n = 3). ( F ) Protein expression levels of CaMKIIα in neurons. n = 3. (○) represents the individual samples within each group. Quantitative results were normalized, and the data in the bar charts are presented as mean ± SD. ** p < 0.01 or *** p < 0.001, **** p < 0.0001.

Article Snippet: The membranes were blocked for 1 h at room temperature with 5% skim milk (Beyotime, Shanghai, China, P0216-1500 g) and incubated overnight at 4 °C with specific primary antibodies, including Cav3.1 (1:2000, NBP2-59322, Novus, Lone Tree, CO, USA), CaMKIIα (1:2000, 3357S, CST, Danvers, MA, USA), CREB (1:2000, 12208-1-AP, Proteintech, Rosemont, IL, USA), BDNF (1:1000, ab226843, abcam, Cambridge, UK), PSD95 (1:1000, 2507S, CST), SYN (1:2000, 17785-1-AP, Proteintech), ProBDNF (1:1000, 28205-1-AP, Proteintech), and GAPDH (1:2000, UM4002, YouKang, Tianjin, China).

Techniques: Control, Western Blot, Expressing