Journal: Frontiers in Endocrinology

Article Title: Identification of immune-related endoplasmic reticulum stress genes in proliferative diabetic retinopathy using bioinformatics analysis

doi: 10.3389/fendo.2024.1341206

Figure Lengend Snippet: External validation of TRAM1, TXNIP and ER stress-related hub genes (A) Validation of ER stress-related hub genes in the GSE60436 dataset. (B) The protein levels of TRAM1 and TXNIP were evaluated in cell samples by western blot. (C) The mRNA levels of BCL2, CCL2, IL-1Β, TLR4, TNF, and TP53 were measured in cell samples by qRT-PCR. Ctrl, control group; HG, high-glucose group. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

Article Snippet: The membranes were then blocked in QuickBlockTM Western Blocking Buffer (P0252, Beyotime) for 30 minutes and incubated overnight at 4°C with primary antibodies against TRAM1 (18243-1-AP, Proteintech) and TXNIP (12705-1-AP, Proteintech).

Techniques: Biomarker Discovery, Western Blot, Quantitative RT-PCR, Control

Journal: Scientific Reports

Article Title: Potential therapeutic effects of IL28RA inhibition on acute myocardial infarction through phosphorylated JAK1/STAT1 signaling pathways

doi: 10.1038/s41598-024-83668-x

Figure Lengend Snippet: This study investigates the relationship between IL28A and IL28B levels, clinical indicators, and the timing of myocardial infarction. It includes a comparison of IL28A and IL28B levels in patients who have experienced myocardial infarction versus control subjects ( A ). Additionally, it examines the correlation between IL28A and IL28B levels and various clinical parameters, including BMI, HDL, cTnT, cTnI, and CK-MB ( B ). Furthermore, the study explores the temporal changes in IL28A and IL28B levels in relation to the progression of myocardial infarction ( C ) ( * P < 0.05, ** P < 0.01, **** P < 0.0001).

Article Snippet: Subsequently, the membrane was blocked with QuickBlockTM Western (Beyotime, p0252) for 20 min at 4 °C, and incubated overnight at 4 °C with primary antibodies against IL28RA (PA5-98608, Invitrogen, USA), IL28A (YT5305, ImmunoWay, USA), IL28B (PA5-103439, Invitrogen, USA), JAK1 (3344P, Cell Signaling Technology, USA), p-JAK1 (3331S, Cell Signaling Technology, USA), STAT1 (9172L, Cell Signaling Technology, USA), p-STAT1 (9167S, Cell Signaling Technology, USA), BAX(50599-2-lg, Proteintech, China) and Bcl2(26593-1-AP, Proteintech, China) at a dilution of 1:2000.

Techniques: Comparison, Control

Journal: Scientific Reports

Article Title: Potential therapeutic effects of IL28RA inhibition on acute myocardial infarction through phosphorylated JAK1/STAT1 signaling pathways

doi: 10.1038/s41598-024-83668-x

Figure Lengend Snippet: Immunofluorescence images of IL28A, IL28B, and IL28RA expression in mouse heart tissue were presented. In comparison to the control group, the expression levels of IL28RA and IL28A were significantly elevated in the AMI group. Conversely, the expression levels of IL28RA and IL28A in the AMI + shIL28RA group were significantly reduced when compared to the AMI group. Importantly, no significant differences in IL28B expression were observed among the groups. ( * P < 0.05, *** P < 0.001, **** P < 0.0001).

Article Snippet: Subsequently, the membrane was blocked with QuickBlockTM Western (Beyotime, p0252) for 20 min at 4 °C, and incubated overnight at 4 °C with primary antibodies against IL28RA (PA5-98608, Invitrogen, USA), IL28A (YT5305, ImmunoWay, USA), IL28B (PA5-103439, Invitrogen, USA), JAK1 (3344P, Cell Signaling Technology, USA), p-JAK1 (3331S, Cell Signaling Technology, USA), STAT1 (9172L, Cell Signaling Technology, USA), p-STAT1 (9167S, Cell Signaling Technology, USA), BAX(50599-2-lg, Proteintech, China) and Bcl2(26593-1-AP, Proteintech, China) at a dilution of 1:2000.

Techniques: Immunofluorescence, Expressing, Comparison, Control

Journal: Scientific Reports

Article Title: Potential therapeutic effects of IL28RA inhibition on acute myocardial infarction through phosphorylated JAK1/STAT1 signaling pathways

doi: 10.1038/s41598-024-83668-x

Figure Lengend Snippet: Western blot analysis demonstrated the expression levels of JAK1, p-JAK1, STAT1, p-STAT1, IL28A, IL28B, and IL28RA in mouse heart tissue. Compared to the control group, the expression levels of p-JAK1/STAT1, p-STAT1/STAT1, IL28A, and IL28RA were significantly elevated in the AMI group. In contrast, the expression levels of p-JAK1/STAT1, p-STAT1/STAT1, IL28A, and IL28RA were significantly reduced in the AMI + shIL28RA group compared to the AMI group. No significant differences were observed in IL28B between the groups. ( * P < 0.05, ** P < 0.01, **** P < 0.0001).

Article Snippet: Subsequently, the membrane was blocked with QuickBlockTM Western (Beyotime, p0252) for 20 min at 4 °C, and incubated overnight at 4 °C with primary antibodies against IL28RA (PA5-98608, Invitrogen, USA), IL28A (YT5305, ImmunoWay, USA), IL28B (PA5-103439, Invitrogen, USA), JAK1 (3344P, Cell Signaling Technology, USA), p-JAK1 (3331S, Cell Signaling Technology, USA), STAT1 (9172L, Cell Signaling Technology, USA), p-STAT1 (9167S, Cell Signaling Technology, USA), BAX(50599-2-lg, Proteintech, China) and Bcl2(26593-1-AP, Proteintech, China) at a dilution of 1:2000.

Techniques: Western Blot, Expressing, Control

Journal: Scientific Reports

Article Title: Potential therapeutic effects of IL28RA inhibition on acute myocardial infarction through phosphorylated JAK1/STAT1 signaling pathways

doi: 10.1038/s41598-024-83668-x

Figure Lengend Snippet: Graphic summary. Following acute myocardial infarction, the level of IL28A in mouse myocardial tissue increased significantly, while the level of IL28B did not show a significant increase. Concurrently, the level of IL28RA also significantly increased, leading to phosphorylation activation of the JAK1/STAT1 pathway and subsequent apoptosis induction. Knocking down IL28RA reversed these changes.

Article Snippet: Subsequently, the membrane was blocked with QuickBlockTM Western (Beyotime, p0252) for 20 min at 4 °C, and incubated overnight at 4 °C with primary antibodies against IL28RA (PA5-98608, Invitrogen, USA), IL28A (YT5305, ImmunoWay, USA), IL28B (PA5-103439, Invitrogen, USA), JAK1 (3344P, Cell Signaling Technology, USA), p-JAK1 (3331S, Cell Signaling Technology, USA), STAT1 (9172L, Cell Signaling Technology, USA), p-STAT1 (9167S, Cell Signaling Technology, USA), BAX(50599-2-lg, Proteintech, China) and Bcl2(26593-1-AP, Proteintech, China) at a dilution of 1:2000.

Techniques: Activation Assay

Journal: Microbiome

Article Title: Xylanase enhances gut microbiota-derived butyrate to exert immune-protective effects in a histone deacetylase-dependent manner.

doi: 10.1186/s40168-024-01934-6

Figure Lengend Snippet: Fig. 8 Butyrate promotes il-17d expression mainly by HDAC inhibitor. A, B, C, D The effects of FFAR2 inhibitor, FFAR3 inhibitor, NAM, TSA, and ITSA on the il-17d expression of intestinal epithelial cells, E the effects of sodium butyrate on the pan-acetylation level of intestinal epithelial cells, F the protein expression of HDAC3, G the effects of RGFP966 (HDAC3 inhibitor) on the il-17d expression of intestinal epithelial cells, and H luciferase reporter plasmid pGL 4.10- il-17d -luc were transiently transfected into 293 T cells. Cells were treated with butyrate or RGFP966 for 12 h. Luciferase activities of untreated cells and butyrate or RGFP966-treated cells are depicted relative to the Renilla activities. Data was expressed as mean ± SEM (n = 3). SB, cells treated with sodium butyrate. FFAR2/3, free fatty acid receptor 2/3. NAM, nicotinamide. TSA, trichostatin A. ITSA, inhibitor of trichostatin. Statistically significant results were expressed by lowercase letters (a, b, c) based on ANOVA with Tukey test

Article Snippet: After blocked with QuickBlockTM Western (P0252, Beyotime Biotechnology, China) for 20 min, the membranes were incubated with the primary antibodies pan-acetylation (66,289–1-Ig, Proteintech, Rosemont, USA), HDAC3 (10,255–1-AP, Proteintech, Rosemont, USA), and α-tubulin (M1501-180, HUABIO, Hangzhou, China) overnight at 4 °C.

Techniques: Expressing, Luciferase, Plasmid Preparation, Transfection

Journal: Microbiome

Article Title: Xylanase enhances gut microbiota-derived butyrate to exert immune-protective effects in a histone deacetylase-dependent manner.

doi: 10.1186/s40168-024-01934-6

Figure Lengend Snippet: Fig. 8 Butyrate promotes il-17d expression mainly by HDAC inhibitor. A, B, C, D The effects of FFAR2 inhibitor, FFAR3 inhibitor, NAM, TSA, and ITSA on the il-17d expression of intestinal epithelial cells, E the effects of sodium butyrate on the pan-acetylation level of intestinal epithelial cells, F the protein expression of HDAC3, G the effects of RGFP966 (HDAC3 inhibitor) on the il-17d expression of intestinal epithelial cells, and H luciferase reporter plasmid pGL 4.10- il-17d -luc were transiently transfected into 293 T cells. Cells were treated with butyrate or RGFP966 for 12 h. Luciferase activities of untreated cells and butyrate or RGFP966-treated cells are depicted relative to the Renilla activities. Data was expressed as mean ± SEM (n = 3). SB, cells treated with sodium butyrate. FFAR2/3, free fatty acid receptor 2/3. NAM, nicotinamide. TSA, trichostatin A. ITSA, inhibitor of trichostatin. Statistically significant results were expressed by lowercase letters (a, b, c) based on ANOVA with Tukey test

Article Snippet: After blocked with QuickBlockTM Western (P0252, Beyotime Biotechnology, China) for 20 min, the membranes were incubated with the primary antibodies pan-acetylation (66,289–1-Ig, Proteintech, Rosemont, USA), HDAC3 (10,255–1-AP, Proteintech, Rosemont, USA), and α-tubulin (M1501-180, HUABIO, Hangzhou, China) overnight at 4 °C.

Techniques: Expressing, Luciferase, Plasmid Preparation, Transfection

Journal: International Journal of Molecular Sciences

Article Title: Effects of MICU1-Mediated Mitochondrial Calcium Uptake on Energy Metabolism and Quality of Vitrified-Thawed Mouse Metaphase II Oocytes

doi: 10.3390/ijms23158629

Figure Lengend Snippet: Effect of vitrification on the mRNA of MICU1 ( A ), the protein of MICU1 ( B , C ) and MCU ( B , D ) expressions in oocytes. Data are presented as the mean ± SEM. of 3 to 5 samples per group. * p < 0.05, ** p < 0.01. Con, control; Tox, toxicity; Vit, vitrified.

Article Snippet: The membranes were blocked with QuickBlockTM Western Blocking Buffer (P0252, Beyotime, Shanghai, China) and incubated with anti-MICU1 mouse monoclonal primary antibody (YM3430, Immunoway, Berkeley, CA, USA), anti-MCU rabbit polyclonal primary antibody (26312-1-AP, Proteintech, Rosemont, Chicago, IL, USA), anti-PDH rabbit monoclonal primary antibody (ab168379, Abcam, Cambridge, UK), anti-PDH (phospho S293) rabbit monoclonal primary antibody (ab177461, Abcam, Cambridge, UK) overnight at 4 °C.

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: Effects of MICU1-Mediated Mitochondrial Calcium Uptake on Energy Metabolism and Quality of Vitrified-Thawed Mouse Metaphase II Oocytes

doi: 10.3390/ijms23158629

Figure Lengend Snippet: MICU1 stimulates mitochondrial Ca 2+ uptake in mice vitrified oocytes. ( A , B ) Analyses of MICU1 protein expression in vitrified oocytes treated with or without MCU-i4. ( C , D ) Analyses of MICU1 protein expression in vitrified oocytes treated with or without DS16570511. ( E ) Fluorescence images showing the mitochondrial Ca 2+ levels in each group. Scale bar = 20 μm. ( F ) Relative fluorescence intensity for mitochondrial Ca 2+ in each group. Data in B, D, F are presented as the mean ± SEM. of 3 to 5 independent experiments per group. ns, no significant, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Con, control; Vit, vitrified; Vit+i4, vitrified+MCU-i4; Vit+ds, vitrified+DS16570511.

Article Snippet: The membranes were blocked with QuickBlockTM Western Blocking Buffer (P0252, Beyotime, Shanghai, China) and incubated with anti-MICU1 mouse monoclonal primary antibody (YM3430, Immunoway, Berkeley, CA, USA), anti-MCU rabbit polyclonal primary antibody (26312-1-AP, Proteintech, Rosemont, Chicago, IL, USA), anti-PDH rabbit monoclonal primary antibody (ab168379, Abcam, Cambridge, UK), anti-PDH (phospho S293) rabbit monoclonal primary antibody (ab177461, Abcam, Cambridge, UK) overnight at 4 °C.

Techniques: Expressing, Fluorescence, Control

Journal: International Journal of Molecular Sciences

Article Title: Effects of MICU1-Mediated Mitochondrial Calcium Uptake on Energy Metabolism and Quality of Vitrified-Thawed Mouse Metaphase II Oocytes

doi: 10.3390/ijms23158629

Figure Lengend Snippet: Sequence and annealing temperature of flfluorescence quantitative PCR primers used. AT, annealing temperature.

Article Snippet: The membranes were blocked with QuickBlockTM Western Blocking Buffer (P0252, Beyotime, Shanghai, China) and incubated with anti-MICU1 mouse monoclonal primary antibody (YM3430, Immunoway, Berkeley, CA, USA), anti-MCU rabbit polyclonal primary antibody (26312-1-AP, Proteintech, Rosemont, Chicago, IL, USA), anti-PDH rabbit monoclonal primary antibody (ab168379, Abcam, Cambridge, UK), anti-PDH (phospho S293) rabbit monoclonal primary antibody (ab177461, Abcam, Cambridge, UK) overnight at 4 °C.

Techniques: Sequencing, Real-time Polymerase Chain Reaction