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 activate UPR signaling to induce ERS . A HD11 cells were treated with 50–200 µg/mL OMVs for 6 h, and the relative mRNA levels of UPR downstream genes ATF4 , CHOP , EDEM1 , ERdj4 , and ATF6 were detected by qPCR. ( n = 3) B HD11 cells were treated with OMVs (100 µg/mL) for 0–9 h, and qPCR was used to detect the relative mRNA levels of UPR downstream genes ATF4 , CHOP , EDEM1 , ERdj4 , and ATF6 . ( n = 3) C , D HD11 cells were treated with OMVs (100 µg/mL) for 0–9 h, western blotting was used to detect the expression of PERK, p-PERK, eIF2α, p-eIF2α, CHOP, IRE1, p-IRE1, and ATF6 proteins in cell lysates, and gray value analysis was performed using ImageJ software ( D ). ( n = 3). E , F HD11 cells were treated with OMVs (100 µg/mL) for 6 h in the absence or presence of 4-PBA (2 mM), western blot analysis was used to detect the expression of PERK, p-PERK, eIF2α, p-eIF2α, CHOP, IRE1, p-IRE1, and ATF6 proteins in cell lysates, and gray value analysis was performed using ImageJ software ( F ). ( n = 3); n represents three biological replicates; data points indicate independent culture systems. Bar charts display mean ± standard error of the mean (SEM). Two-way ANOVA with Sidak correction was performed (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Rabbit anti-ATF6 polyclonal antibody , Boster, Hubei, China , 1:2000.

Techniques: Western Blot, Expressing, 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-ATF6 polyclonal antibody , Boster, Hubei, China , 1:2000.

Techniques: Infection, Expressing, Activation Assay

Journal: Virologica Sinica

Article Title: Modulation of the unfolded protein response by white spot syndrome virus via wsv406 targeting BiP to facilitate viral replication

doi: 10.1016/j.virs.2024.10.005

Figure Lengend Snippet: Wsv406 doesn't interfere the interaction of LvBiP with ATF6, IRE1 and PERK, but suppresses LvBiP ATPase activity. A His-tag pulldown for the detection of the interaction between BiP and ATF6, IRE1 or PERK by expression of wsv406 in a dose-dependent manner. The results were shown via western blotting. B Schematic representation of the procedures for measuring the ATPase activity of BiP. C ATPase activity of the recombinant BiP. GST-BiP were bound to Glutathione beads and MBP-wsv406 or MBP and then assessed for the ATPase activity. The values were normalized using the measure of empty beads.

Article Snippet: The following steps were the same as above, the primary antibodies used were rabbit anti-ATF4 antibody (Bioss, bs-1531R), rabbit anti-ATF6 antibody (Bioss, bs-1634R) and mouse anti-β-actin antibody (Sigma, A1978).

Techniques: Activity Assay, Expressing, Western Blot, Recombinant

Journal: Virologica Sinica

Article Title: Modulation of the unfolded protein response by white spot syndrome virus via wsv406 targeting BiP to facilitate viral replication

doi: 10.1016/j.virs.2024.10.005

Figure Lengend Snippet: Effects of wsv406 knockdown on ATF6 pathway during WSSV infection. A Transcription factor LvATF6 nuclear translocation in wsv406 silenced hemocytes infected by WSSV. The percentage of ATF6 nuclear translocation was calculated by Image J. The scale bar ​= ​5 ​μm. B–D Expression levels of LvASK1 ( B ), LvXBP1 ( C ) and WSSV genes ( D ) in hemocytes and gills of dsRNA-GFP treated or dsRNA-wsv406 treated shrimps at 48 ​h post WSSV infection. Differences were analyzed using Student's t -test ( ∗∗ P ​< ​0.01).

Article Snippet: The following steps were the same as above, the primary antibodies used were rabbit anti-ATF4 antibody (Bioss, bs-1531R), rabbit anti-ATF6 antibody (Bioss, bs-1634R) and mouse anti-β-actin antibody (Sigma, A1978).

Techniques: Knockdown, Infection, Translocation Assay, Expressing

Journal: Virologica Sinica

Article Title: Modulation of the unfolded protein response by white spot syndrome virus via wsv406 targeting BiP to facilitate viral replication

doi: 10.1016/j.virs.2024.10.005

Figure Lengend Snippet: The possible model illustrating that WSSV hijacks UPR signaling through wsv406 targeting BiP that facilitates to WSSV replication. In WSSV-infected shrimp, the viral protein wsv406 interacts with and inhibits the ATPase activity of the host protein BiP. This interaction leads to the activation of the PERK-eIF2α and ATF6 pathways, which subsequently promote the expression of other viral genes, facilitating WSSV replication.

Article Snippet: The following steps were the same as above, the primary antibodies used were rabbit anti-ATF4 antibody (Bioss, bs-1531R), rabbit anti-ATF6 antibody (Bioss, bs-1634R) and mouse anti-β-actin antibody (Sigma, A1978).

Techniques: Infection, Activity Assay, Activation Assay, Expressing