Journal: Journal of Advanced Research

Article Title: Tankyrase activity is essential for asymmetric division and chromosome segregation in oocyte meiosis

doi: 10.1016/j.jare.2025.07.008

Figure Lengend Snippet: TNKS affects distributions of Formin2 and ER for chromosome migration in mouse oocytes. (A) Representative images of Formin2 in MI stage oocyte from the control and JW55 treatment groups. Green, Formin2. Cyan, DNA. Bar = 20 μm. (B) Relative intensity of Formin2 at spindle poles in oocytes from the control (n = 47) and JW55 treatment (n = 51) groups. ***, P < 0.001. (C) Band intensity analysis of Formin2 expression in the MI stage oocytes from control and JW55 treatment groups. *, P < 0.05. (D) Representative images of ER-tracker in MI stage oocyte from the control and JW55 treatment groups. Black, ER-tracker. Bar = 20 μm. (E) The percentage of abnormal ER distribution in oocytes from the control (n = 50) and JW55-treated (n = 56) groups. *, P < 0.05. (F) Relative intensity of ER-tracker in oocytes from the control (n = 50) and JW55-treated (n = 56) groups. ***, P < 0.001. (G) Western blot of GRP78 in MI stage oocytes from control and JW55 treatment groups. (H) Band intensity analysis of GRP78 in the control and JW55 treatment groups. *, P < 0.05. (I) Representative images of a relative position change between chromosomes and Formin2 in ATI stage oocytes from the control and JW55 treatment groups. Green, Formin2. Cyan, DNA. Bar = 20 μm. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Rabbit anti-α-Tubulin antibody (11224-1-AP), rabbit polyclonal anti-GAPDH antibody (10494-1-AP), rabbit anti-GRP78 antibody (11587-1-AP), rabbit anti-TNKS1 antibody (18030-1-AP), rabbit anti-Ran antibody (10469-1-AP), and rabbit anti-Rab11a antibody (67902-1-Ig), rabbit anti-Securin antibody (18040-1-AP), rabbit anti-CDC25C antibody (66912-1-Ig), rabbit anti-Formin2 antibody (11259-1-AP), rabbit anti-BubR1 (11504-2-AP) were from Proteintech.

Techniques: Migration, Control, Expressing, Western Blot

Journal: International Dental Journal

Article Title: Baicalin Alleviates LPS-Induced Apoptosis of Periodontal Ligament Cells Via Inhibiting Endoplasmic Reticulum Stress

doi: 10.1016/j.identj.2025.109365

Figure Lengend Snippet: BA attenuates Pg -LPS-induced ER stress in hPDLCs. hPDLCs were pretreated with varying concentrations of BA and subsequently exposed to Pg -LPS (1 µg/mL) for 24h to assess its impact on ER stress. A, Schematic diagram illustrating the experimental workflow. B, Heatmap representing the relative mRNA expression levels of ER stress-related genes including GRP78, PERK, ATF6, and IRE1α, as measured by quantitative RT-PCR. C, ER-Tracker staining was performed to visualize and assess ER integrity and stress levels. Representative fluorescence images show ER morphology and signal intensity under different treatment conditions. D-H, Western blot analysis and quantification of ER stress-related proteins, including GRP78, PERK, IRE1α, and ATF6. Data are expressed as mean ± SD, n = 3. * P < .05, ** P < .01, *** P < .001, **** P < .0001.

Article Snippet: After blocking with 5% nonfat milk, membranes were incubated overnight at 4 °C with primary antibodies against GRP78 (CST, #3177), PERK (CST, #5683), ATF6 (CST, #65880), IRE1α (CST, #3294), eIF2α (CST, #9722), p-eIF2α (CST, #3398), ATF4 (Huabio, ET1612-37), CHOP (CST, #2895), Bcl-2 (Abmart, T40056 ), BAX (Abmart, T40051 ), and Caspase-3 (Huabio, SU38-04).

Techniques: Expressing, Quantitative RT-PCR, Staining, Fluorescence, Western Blot

Journal: Veterinary Research

Article Title: Outer membrane vesicles secreted by avian pathogenic Escherichia coli promote its survival within macrophages and systemic infection by inducing endoplasmic reticulum stress-mediated autophagy flux blockade

doi: 10.1186/s13567-025-01679-6

Figure Lengend Snippet: APEC OMVs induce ERS in HD11 cells to evade immune clearance by macrophages . Transmission electron microscopy shows ER network expansion and swelling in HD11 cells treated with WT, WTΔ ypjA , or OMVs. The data represent one of three independent experiments. Scale bar, 2 μm. B OMVs colocalized with ER. DiO-OMVs were incubated with HD11 cells at 37 °C for 6 h, the ER was labeled with ER-Tracker Red, and laser confocal microscopy imaging was performed. The data represent one of three independent experiments. Scale bar, 7.5 μm. Colocalization scatter plot was generated using ImageJ software and GraphPad Prism 8. C HD11 cells were treated with 50–200 µg/mL OMVs for 6 h, and the relative mRNA level of GRP78/BiP was determined by qPCR. ( n = 3) D HD11 cells were treated with OMVs (100 µg/mL) for 0–9 h, and the relative mRNA level of GRP78/BiP was determined by qPCR. ( n = 3) E , F HD11 cells were treated with OMVs (100 µg/mL) for 0–9 h, and western blot was used to detect the expression of GRP78/BiP protein in cell lysates and gray value analysis was performed using ImageJ software ( F ). ( n = 3) G Cell viability of HD11 cells treated with OMVs (200 µg/mL) for 12 h was detected by CCK-8 in the absence or presence of 4-PBA (2 mM). ( n = 3) H , I Intracellular survival of HD11 cells infected with WT at MOI = 100 in the absence or presence of 4-PBA (2 mM). Square culture plates showed 1/ 100 of the bacterial load. ( n = 3). n represents three biological replicates; data points indicate independent culture systems. Bar charts display mean ± standard error of the mean (SEM). Data were analyzed using Student’s t -test and two-way ANOVA with Sidak correction. (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Rabbit anti-GRP78/BiP polyclonal antibody , Abmart, Shanghai, China , 1:5000.

Techniques: Transmission Assay, Electron Microscopy, Incubation, Labeling, Confocal Microscopy, Imaging, Generated, Software, Western Blot, Expressing, CCK-8 Assay, Infection

Journal: Veterinary Research

Article Title: Outer membrane vesicles secreted by avian pathogenic Escherichia coli promote its survival within macrophages and systemic infection by inducing endoplasmic reticulum stress-mediated autophagy flux blockade

doi: 10.1186/s13567-025-01679-6

Figure Lengend Snippet: APEC OMVs induce ERS to promote APEC systemic infection . Flowchart of the infection experiment using chicks treated with 4-PBA. B On the sixth day after WT infection of chicks (1 × 10 CFU/chick), the number of bacteria (CFU) was counted in the trachea, lungs, liver, and spleen tissues of the chicks in the absence or presence of 4-PBA (50 mg/kg). Trachea: 95% CI [0.26–0.38]; lungs: 95% CI [0.65–0.78]; liver: 95% CI [0.004–0.13]; spleen: 95% CI [0.08–0.21]. C On the sixth day after WT infection of chicks (1 × 10 9 CFU/chick), HE staining was performed on tracheal, lung, liver, and spleen tissues of the chicks in the absence or presence of 4-PBA (50 mg/kg). Trachea: degeneration and hyperplasia of epithelial mucosal cells (black arrow), diffuse neutrophilic infiltration (red arrow), mucus and inflammatory exudate in the mucosa of the tracheal wall (blue arrow); lung: markedly widened pulmonary septa (black arrow), inflammatory cell infiltration in the pulmonary interstitium (red arrow); liver: hepatocytes show lytic degeneration (black arrow) with inflammatory cell infiltration (red arrow); spleen: marked hyperplasia of lymphoid follicles (black arrow), blurred demarcation between white and red pulp (red arrow). Scale bar, 50 µm or 200 µm. D – F GRP78/BiP immunofluorescence staining was performed on lung and spleen tissues from chicks on day 6 post-infection ( D ) and analyzed using ImageJ software ( E , F ). Scale bar, 50 µm. Data points represent independent cultures; bar charts show mean ± standard error of the mean (SEM). Two-way ANOVA with Sidak correction was performed (* p < 0.05, ** p < 0.01, *** p < 0.001). Effect sizes with 95% confidence intervals are reported in panel ( B ).

Article Snippet: Rabbit anti-GRP78/BiP polyclonal antibody , Abmart, Shanghai, China , 1:5000.

Techniques: Infection, Bacteria, Staining, Immunofluorescence, Software

Journal: Veterinary Research

Article Title: Outer membrane vesicles secreted by avian pathogenic Escherichia coli promote its survival within macrophages and systemic infection by inducing endoplasmic reticulum stress-mediated autophagy flux blockade

doi: 10.1186/s13567-025-01679-6

Figure Lengend Snippet: OMVs secreted by avian pathogenic Escherichia coli promote its survival within macrophages and systemic infection by inducing ERS-mediated autophagy flux blockade . APEC-secreted OMVs, upon uptake by HD11 cells, induce ROS accumulation and Ca 2+ release, triggering ERS and activating UPR pathways, including the PERK, IRE1, and ATF6 signaling branches, leading to the expression of stress-related factors such as GRP78/BiP and CHOP. The sustained activation of ERS inhibits autophagosome degradation and disrupts the acidic environment of lysosomes, thereby preventing autophagosomes from fusing with lysosomes and impairing the phagocytic clearance capacity of macrophages. Collectively, these abnormal conditions facilitate APEC survival within HD11 cells and enable immune evasion, ultimately promoting bacterial dissemination and systemic infection in the host.

Article Snippet: Rabbit anti-GRP78/BiP polyclonal antibody , Abmart, Shanghai, China , 1:5000.

Techniques: Infection, Expressing, Activation Assay

Journal: Scientific Reports

Article Title: A pro-inflammatory neutrophil subpopulation drives intestinal ischemia–reperfusion injury via the ATF4-mediated endoplasmic reticulum stress pathway

doi: 10.1038/s41598-026-36938-9

Figure Lengend Snippet: Pharmacological modulation of ER stress influences I/R injury severity. ( A ) The expression level of serum DAO in mice was determined using an ELISA kit. ( B ) Serum IL-6 expression in mice was assessed by ELISA. ( C, D ) Quantification of small intestinal length in mice. ( E, F ) H&E staining of mouse small intestine and histological evaluation using Chiu’s scoring system. ( G-I ) Immunofluorescence staining of mouse small intestine for Ly6G and ATF4 expression. ( J-M ) Expression levels of the intestinal mucosal barrier protein Occludin and endoplasmic reticulum stress markers ATF4 and GRP78 in mouse intestinal tissue. Data are presented as mean ± SD. Statistical significance is indicated as *P < 0.05, **P < 0.01, ***P < 0.001, **P < 0.0001. NC, normal control; IR, ischemia–reperfusion; Tun, tunicamycin; KO, knockout; neu, neutrophils.

Article Snippet: After blocking (Servicebio), membranes were incubated with primary antibodies against ATF4 (Proteintech, 10,835–1-AP; 1:1000), GRP78 (CST, 3177 T; 1:1000), Occludin (CST, 91131 T; 1:1000), and β-actin (CST, 4970 T; 1:3000).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Staining, Immunofluorescence, Control, Knock-Out

Journal: Scientific Reports

Article Title: A pro-inflammatory neutrophil subpopulation drives intestinal ischemia–reperfusion injury via the ATF4-mediated endoplasmic reticulum stress pathway

doi: 10.1038/s41598-026-36938-9

Figure Lengend Snippet: Neutrophils induce MODE-K cell injury through endoplasmic reticulum stress. ( A ) IL-6 expression in co-culture supernatant was quantified using an ELISA kit. ( B ) Cell viability of MODE-K cells was assessed using the CCK-8 assay. ( C ) Transmission electron microscopy (TEM) of MODE-K cells. ( D, E ) Apoptosis in MODE-K cells was detected by TUNEL staining. ( F-I ) Expression levels of the tight junction protein Occludin and endoplasmic reticulum stress markers ATF4 and GRP78 in MODE-K cells. Data are presented as mean ± SD. Statistical significance is indicated as *P < 0.05, **P < 0.01, ***P < 0.001, **P < 0.0001. NC, normal control; IR, ischemia–reperfusion; Tun, tunicamycin; KO, knockout; neu, neutrophils.

Article Snippet: After blocking (Servicebio), membranes were incubated with primary antibodies against ATF4 (Proteintech, 10,835–1-AP; 1:1000), GRP78 (CST, 3177 T; 1:1000), Occludin (CST, 91131 T; 1:1000), and β-actin (CST, 4970 T; 1:3000).

Techniques: Expressing, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Transmission Assay, Electron Microscopy, TUNEL Assay, Staining, Control, Knock-Out