Journal: bioRxiv

Article Title: Chemical modulation of the unfolded protein response reveals an antiviral role for the PERK pathway in human coronavirus 229E infection

doi: 10.1101/2025.09.11.675615

Figure Lengend Snippet: (A) A549 cells were treated with DMSO, KIRA6 (10 μM), or Ceapin-A7 (6 μM) for 24 hours. Alamar blue assay was used to assess cell viability based on cellular metabolic activity. (B-C) A549 cells were primed with DMSO or Tg (0.05 μM) in the absence or presence of KIRA6 (10 μM) or Ceapin-A7 (6 μM). Cell lysates were collected at 24 hours post-treatment. Xbp1s (B) or HERPUD1 (C) protein expression was assessed by western blot. (D-F) A549 cells were primed with DMSO or Tg (0.05 μM) in the presence or absence of KIRA6 (10 μM), prior to infection with HCoV-229E (MOI 0.05). Viral inoculum was removed 2 hours later and replaced with media containing DMSO or KIRA6 (10 μM). (D-F) Cell lysates were collected at 24 hpi to assess XBP1s and N gene expression by RT-qPCR (D, F) , or Xbp1s and HCoV-229E N protein expression by western blot (E) . (G-I) A549 cells were primed with DMSO or Tg (0.05 μM) in the presence or absence of Ceapin-A7 (6 μM) prior to infection with HCoV-229E (MOI 0.05). Viral inoculum was removed 2 hours later and replaced with media containing DMSO or Ceapin-A7 (6 μM). (G-I) Cell lysates were collected at 24 hpi to assess HERPUD1 and N by RT-qPCR (G, I) , or HERPUD1 and HCoV-229E N protein expression by western blot (H) . RT-qPCR data are normalized to actin and set relative to DMSO. (J) A549 cells were primed with DMSO or Tg (0.05 μM) in the presence or absence of KIRA6 (10 μM) or Ceapin-A7 (6 μM), alone or in combination, prior to infection with HCoV-229E (MOI 0.05). Inoculum was removed 2 hours later and replaced with media containing the respective inhibitor(s). Viral supernatants were collected 24 hpi and titrated by plaque assay on Huh7 cells. Graphs show mean +/− SEM from three independent experiments performed in triplicate (*p<0.05, ****p<0.0001). Representative western blots are shown.

Article Snippet: Primary antibodies against IRE1α (Cell Signaling Technology (CST) 3294, dilution 1:1000), ATF6 (CST 8089, dilution 1:1000), PERK (CST 5683, dilution 1:1000), XBP1s (CST 12782T, dilution 1:1000), HERPUD1 (Abcam ab150424, dilution 1:1000), eIF2α-P (Ser-51) (CST 3398S, dilution 1:1000), HCoV-229E N (SinoBiological 40640-T62, dilution 1:10,000), beta-actin (Abcam ab8226, dilution 1:5000), alpha-tubulin (Thermo Scientific MA5-31466, dilution 1:5000), and GAPDH (Thermo Scientific MA5-15738, dilution 1:1000) were used for western blotting.

Techniques: Alamar Blue Assay, Activity Assay, Expressing, Western Blot, Infection, Gene Expression, Quantitative RT-PCR, Plaque Assay

Journal: European Journal of Histochemistry : EJH

Article Title: SRSF3 promotes the generation of XBP1s to stabilize autophagy and enhance hypoxia adaptation in glioma

doi: 10.4081/ejh.2026.4530

Figure Lengend Snippet: Hypoxic glioma cells exhibit increased cell death accompanied by elevated endoplasmic reticulum stress levels and blocked autophagic flux. A ) Western blot analysis of HIF-1α expression in U87 cells. B ) LDH release assay to detect cell death. C ) Western blot analysis of cleaved-caspase3 expression in U87 cells. D ) Western blot analysis of GRP78, CHOP, and ATF4 expression in U87 cells. E ) RT-PCR detection of XBP1u and XBP1s expression. F ) Western blot analysis of XBP1u and XBP1s expression in U87 cells. G ) Western blot analysis of LC3-II and p62 expression in U87 cells. H ) mRFP-GFP-LC3 staining to assess autophagic flux. n=3 independent experiments; * p <0.05, ** p <0.01, *** p <0.001.

Article Snippet: The primary antibodies used were against the following: SRSF3 (1:1000; ab198291; Abcam, Cambridge, UK), HIF-1α (1:1000; ab179483; Abcam), GRP78 (1:2000; 11587-1-AP; Proteintech, Rosemont, IL, USA), CHOP (1:1000; 15204-1-AP; Proteintech), ATF4 (1:1000; 10835-1-AP; Proteintech), LC3 (1:2000; ab192890; Abcam), p62 (1:1000; ab109012; Abcam), cleaved-caspase3 (1:1000; #9661; Cell Signaling Technology, Inc., Danvers, MA, USA), XBP1u (1:1000; 25997-1-AP; Proteintech), XBP1s (1:1000; 24868-1-AP; Proteintech), and β-actin (1:1000; ab8227; Abcam).

Techniques: Western Blot, Expressing, Lactate Dehydrogenase Assay, Reverse Transcription Polymerase Chain Reaction, Staining

Journal: European Journal of Histochemistry : EJH

Article Title: SRSF3 promotes the generation of XBP1s to stabilize autophagy and enhance hypoxia adaptation in glioma

doi: 10.4081/ejh.2026.4530

Figure Lengend Snippet: SRSF3 promotes XBP1s formation. A ) RT‒PCR detection of XBP1u and XBP1s expression in each group of cells. B ) Western blot detection of XBP1u and XBP1s expression in each group of cells. n=3 independent experiments; * p <0.05, ** p <0.01, *** p <0.001.

Article Snippet: The primary antibodies used were against the following: SRSF3 (1:1000; ab198291; Abcam, Cambridge, UK), HIF-1α (1:1000; ab179483; Abcam), GRP78 (1:2000; 11587-1-AP; Proteintech, Rosemont, IL, USA), CHOP (1:1000; 15204-1-AP; Proteintech), ATF4 (1:1000; 10835-1-AP; Proteintech), LC3 (1:2000; ab192890; Abcam), p62 (1:1000; ab109012; Abcam), cleaved-caspase3 (1:1000; #9661; Cell Signaling Technology, Inc., Danvers, MA, USA), XBP1u (1:1000; 25997-1-AP; Proteintech), XBP1s (1:1000; 24868-1-AP; Proteintech), and β-actin (1:1000; ab8227; Abcam).

Techniques: Expressing, Western Blot

Journal: Heliyon

Article Title: Guilu Erxian glue reduces endoplasmic reticulum stress-mediated apoptosis and restores the balance of extracellular matrix synthesis and degradation in chondrocytes by inhibiting the ATF6/GRP78/CHOP signaling pathway

doi: 10.1016/j.heliyon.2024.e39987

Figure Lengend Snippet: Primer list for quantitative polymerase chain reaction.

Article Snippet: After transfer, the membrane was incubated with a BSA (Bovine Serum Albumin) sealant for approximately 2 h at room temperature.Western blot analysis was performed on the following proteins: CHOP (1:1000, catalog number 15204-1-ap), COL2A1 (1:2000, catalog number 28459-1-ap), XBP1 (1:1000, catalog number 24168-1-ap), MMP-13 (1:2000, catalog number 18165-1-AP), ATF6 (1:3000, catalog number 24169-1-AP), GRP78 (1:2000, catalog number 11587-1-AP), and GAPDH (1:5000, catalog number 60004-1-1G), all sourced from Proteintech (Wuhan, Hubei).

Techniques:

Journal: Heliyon

Article Title: Guilu Erxian glue reduces endoplasmic reticulum stress-mediated apoptosis and restores the balance of extracellular matrix synthesis and degradation in chondrocytes by inhibiting the ATF6/GRP78/CHOP signaling pathway

doi: 10.1016/j.heliyon.2024.e39987

Figure Lengend Snippet: GEG Inhibited of TG-induced ERS in chondrocyte. (A) The qPCR method was used to evaluate XBP1 mRNA expression. (B-C) The Western blot technique was applied to detect XBP1 protein levels. (D-E) Immunofluorescence was employed to assess SEC61 expression.The results represent mean ± SD(n = 3).∗ P < 0.05,Compared with Control group; △ P < 0.05,Compared with the TG group; ▲ P < 0.05,Compared with the GEG group.

Article Snippet: After transfer, the membrane was incubated with a BSA (Bovine Serum Albumin) sealant for approximately 2 h at room temperature.Western blot analysis was performed on the following proteins: CHOP (1:1000, catalog number 15204-1-ap), COL2A1 (1:2000, catalog number 28459-1-ap), XBP1 (1:1000, catalog number 24168-1-ap), MMP-13 (1:2000, catalog number 18165-1-AP), ATF6 (1:3000, catalog number 24169-1-AP), GRP78 (1:2000, catalog number 11587-1-AP), and GAPDH (1:5000, catalog number 60004-1-1G), all sourced from Proteintech (Wuhan, Hubei).

Techniques: Expressing, Western Blot, Immunofluorescence, Control