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anti phospho eif2α ser51  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho eif2α ser51
    SAR405 inhibits autophagy during erythropoiesis (A) RT-qPCR in WT murine splenic erythroblast cells (Lineage − CD71 + ) 3 days after acute hemolytic anemia. N = 3 mice per group. Error bars represent standard error of mean (SEM). ∗∗∗ p < 0.001 (two-tailed unpaired Student’s t test). (B) RT-qPCR in murine splenic erythroblasts (lineage − CD71 + Kit + ) cells isolated from control (WT) or Berkeley Sickle Cell Disease (SCD) mice ( N = 3 per genotype). Error bars represent standard error of mean (SEM). ∗ p < 0.05; ∗∗ p < 0.01 (two-tailed unpaired Student’s t test). Target mRNA expression levels were normalized to 18S rRNA. (C) Western blotting of Lineage − CD71 + cells isolated from control (PBS-treated) and anemia (PHZ-treated) spleens for autophagy-related proteins. N = 3 WT mice per experimental group. (D) Semi-quantitative densitometry of p -Ampk (Thr172) relative to total Ampk and p <t>-eIF2α</t> <t>(Ser51)</t> relative to total eIF2α ( N = 3 mice). Error bars represent standard error of mean (SEM). ∗ p < 0.05; ∗∗∗ p < 0.001 (two-tailed unpaired Student’s t test). (E) Experimental layout of splenic EPCs isolated from Samd14 fl/fl (WT) and Samd14 fl/fl;Vav−Cre (S14-CKO) mice post PHZ-induced anemia. EPCs were retrovirally infected to express the GFP-LC3-RFP probe (Addgene #84573). After 24 h, cells were treated with VPS34 inhibitor SAR405 for an additional 48 h before analysis by flow cytometry. (F) Schematic representation of autophagic flux measurement using GFP-LC3-RFP probe. (G) Gating strategy for GFP and RFP signal measurement by flow cytometry. Progressively maturing erythroid cells (R1-R4) were gated within a viable RFP-expressing cell population using cell surface markers CD71 and TER119. (H) Quantitation of GFP/RFP median fluorescence intensity ratio calculated within R1 (CD71 low/med TER119 - ), R2 (CD71 high TER119 - ), R3 (CD71 high TER119 + ), and R4 (CD71 low/neg TER119 + ) populations in DMSO or SAR405-treated WT cultures ( n = 3 mice). Error bars represent standard error of mean (SEM). ∗∗∗ p < 0.001 (One-way ANOVA). (I) Quantitation of GFP/RFP median fluorescence intensity ratio calculated within R2 (CD71 high TER119 - ) and R4 (CD71 low/neg TER119 + ) populations in DMSO or SAR405-treated WT and S14-CKO cultures ( n = 3 mice per group). Error bars represent standard deviation (SD). ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001 (two-way ANOVA).
    Anti Phospho Eif2α Ser51, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/phospho+eif2%CE%B1/pmc13062518-11-0-7
    Average 86 stars, based on 1 article reviews
    anti phospho eif2α ser51 - by Bioz Stars, 2026-10
    86/100 stars

    Images

    1) Product Images from "A stress-induced PI3P complex controls autophagy in erythroid precursors"

    Article Title: A stress-induced PI3P complex controls autophagy in erythroid precursors

    Journal: iScience

    doi: 10.1016/j.isci.2026.115182

    SAR405 inhibits autophagy during erythropoiesis (A) RT-qPCR in WT murine splenic erythroblast cells (Lineage − CD71 + ) 3 days after acute hemolytic anemia. N = 3 mice per group. Error bars represent standard error of mean (SEM). ∗∗∗ p < 0.001 (two-tailed unpaired Student’s t test). (B) RT-qPCR in murine splenic erythroblasts (lineage − CD71 + Kit + ) cells isolated from control (WT) or Berkeley Sickle Cell Disease (SCD) mice ( N = 3 per genotype). Error bars represent standard error of mean (SEM). ∗ p < 0.05; ∗∗ p < 0.01 (two-tailed unpaired Student’s t test). Target mRNA expression levels were normalized to 18S rRNA. (C) Western blotting of Lineage − CD71 + cells isolated from control (PBS-treated) and anemia (PHZ-treated) spleens for autophagy-related proteins. N = 3 WT mice per experimental group. (D) Semi-quantitative densitometry of p -Ampk (Thr172) relative to total Ampk and p -eIF2α (Ser51) relative to total eIF2α ( N = 3 mice). Error bars represent standard error of mean (SEM). ∗ p < 0.05; ∗∗∗ p < 0.001 (two-tailed unpaired Student’s t test). (E) Experimental layout of splenic EPCs isolated from Samd14 fl/fl (WT) and Samd14 fl/fl;Vav−Cre (S14-CKO) mice post PHZ-induced anemia. EPCs were retrovirally infected to express the GFP-LC3-RFP probe (Addgene #84573). After 24 h, cells were treated with VPS34 inhibitor SAR405 for an additional 48 h before analysis by flow cytometry. (F) Schematic representation of autophagic flux measurement using GFP-LC3-RFP probe. (G) Gating strategy for GFP and RFP signal measurement by flow cytometry. Progressively maturing erythroid cells (R1-R4) were gated within a viable RFP-expressing cell population using cell surface markers CD71 and TER119. (H) Quantitation of GFP/RFP median fluorescence intensity ratio calculated within R1 (CD71 low/med TER119 - ), R2 (CD71 high TER119 - ), R3 (CD71 high TER119 + ), and R4 (CD71 low/neg TER119 + ) populations in DMSO or SAR405-treated WT cultures ( n = 3 mice). Error bars represent standard error of mean (SEM). ∗∗∗ p < 0.001 (One-way ANOVA). (I) Quantitation of GFP/RFP median fluorescence intensity ratio calculated within R2 (CD71 high TER119 - ) and R4 (CD71 low/neg TER119 + ) populations in DMSO or SAR405-treated WT and S14-CKO cultures ( n = 3 mice per group). Error bars represent standard deviation (SD). ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001 (two-way ANOVA).
    Figure Legend Snippet: SAR405 inhibits autophagy during erythropoiesis (A) RT-qPCR in WT murine splenic erythroblast cells (Lineage − CD71 + ) 3 days after acute hemolytic anemia. N = 3 mice per group. Error bars represent standard error of mean (SEM). ∗∗∗ p < 0.001 (two-tailed unpaired Student’s t test). (B) RT-qPCR in murine splenic erythroblasts (lineage − CD71 + Kit + ) cells isolated from control (WT) or Berkeley Sickle Cell Disease (SCD) mice ( N = 3 per genotype). Error bars represent standard error of mean (SEM). ∗ p < 0.05; ∗∗ p < 0.01 (two-tailed unpaired Student’s t test). Target mRNA expression levels were normalized to 18S rRNA. (C) Western blotting of Lineage − CD71 + cells isolated from control (PBS-treated) and anemia (PHZ-treated) spleens for autophagy-related proteins. N = 3 WT mice per experimental group. (D) Semi-quantitative densitometry of p -Ampk (Thr172) relative to total Ampk and p -eIF2α (Ser51) relative to total eIF2α ( N = 3 mice). Error bars represent standard error of mean (SEM). ∗ p < 0.05; ∗∗∗ p < 0.001 (two-tailed unpaired Student’s t test). (E) Experimental layout of splenic EPCs isolated from Samd14 fl/fl (WT) and Samd14 fl/fl;Vav−Cre (S14-CKO) mice post PHZ-induced anemia. EPCs were retrovirally infected to express the GFP-LC3-RFP probe (Addgene #84573). After 24 h, cells were treated with VPS34 inhibitor SAR405 for an additional 48 h before analysis by flow cytometry. (F) Schematic representation of autophagic flux measurement using GFP-LC3-RFP probe. (G) Gating strategy for GFP and RFP signal measurement by flow cytometry. Progressively maturing erythroid cells (R1-R4) were gated within a viable RFP-expressing cell population using cell surface markers CD71 and TER119. (H) Quantitation of GFP/RFP median fluorescence intensity ratio calculated within R1 (CD71 low/med TER119 - ), R2 (CD71 high TER119 - ), R3 (CD71 high TER119 + ), and R4 (CD71 low/neg TER119 + ) populations in DMSO or SAR405-treated WT cultures ( n = 3 mice). Error bars represent standard error of mean (SEM). ∗∗∗ p < 0.001 (One-way ANOVA). (I) Quantitation of GFP/RFP median fluorescence intensity ratio calculated within R2 (CD71 high TER119 - ) and R4 (CD71 low/neg TER119 + ) populations in DMSO or SAR405-treated WT and S14-CKO cultures ( n = 3 mice per group). Error bars represent standard deviation (SD). ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001 (two-way ANOVA).

    Techniques Used: Quantitative RT-PCR, Two Tailed Test, Isolation, Control, Expressing, Western Blot, Infection, Flow Cytometry, Quantitation Assay, Fluorescence, Standard Deviation

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    Article Title: H19 Is a PERK-Regulated Long Non-Coding RNA That Fine-Tunes UPR Signalling and Inhibits Endoplasmic Reticulum Stress-Induced Cell Death.
    Article Snippet: .. The membrane was incubated overnight at 4 ◦C with primary antibodies against ATF6 (Abcam, Cambridge, UK; Cat #ab122897), spliced XBP1 (BioLegend, San Diego, CA, USA; Cat #619502), PERK (Cell Signalling, Danvers, MA, USA; Cat #C33E10), phospho-eIF2α (Cell Signalling Technology, Danvers, MA, USA; Cat #9721S), β-actin (Sigma-Aldrich, St. Louis, MO, USA; Cat #A-5060), HA-Tag (Cell Signaling Technology, Danvers, MA, USA; Cat #MA1-12429), and/or Trimethyl-histone H3 (K4) (Cell Signaling Technology, Danvers, MA, USA; Cat #9727S). .. After three washes with PBST, the membrane was incubated with the 1:5000 diluted horseradish peroxidase-conjugated anti-rabbit IgG (Cell Signaling Technology, Danvers, MA, USA; Cat #7074) or anti-mouse IgG (Cell Signaling Technology, Danvers, MA, USA; Cat #7076) secondary antibody for 2 h at room temperature.

    Incubation:

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    Nucleic Acid Electrophoresis:

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    Western Blot:

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    Saline:

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    A Schematic illustration showing how PC/PE imbalance due to impaired SAM-dependent methylation leads to ER stress and mitochondrial dysfunction. The illustration was created in BioRender. Zang, Y. (2026) https://BioRender.com/juq60sz . B Heatmap of individual PC/PE species ratios from hepatic lipidomic analysis comparing sham-GAN + CCl 4 and VSG-GAN + CCl 4 groups ( n = 9, 10, respectively). C Total hepatic PC/PE ratio in sham-GAN + CCl 4 , and VSG-GAN+CCl 4 groups ( n = 9, 10 respectively). D Western blot analysis of ER stress markers (P-PERK, PERK, <t>P-eIF2α,</t> eIF2α) in liver samples from sham-Chow, sham-GAN+CCl 4 and VSG-GAN + CCl 4 groups ( n = 3 per group). E Densitometric quantification of p-PERK/PERK and p-eIF2α/eIF2α ratios from panel ( D ). F Quantification of mitochondrial ROS (mROS) production in isolated hepatocytes from indicated groups ( n = 3 per group). G Quantification of hepatic malondialdehyde (MDA) levels as a marker of lipid peroxidation ( n = 6, 8, 8 for sham-Chow, sham-GAN + CCl 4 , and VSG-GAN + CCl 4 , respectively). H Quantification of mitochondrial DNA to nuclear DNA ratio indicating mitochondrial content in liver samples ( n = 6, 8, 8 for sham-Chow, sham-GAN + CCl 4 , and VSG-GAN + CCl 4 , respectively). I Quantification of hepatic ATP content ( n = 6, 8, 8 for sham-Chow, sham-GAN + CCl 4 , and VSG-GAN + CCl 4 , respectively). All data are presented as mean ± SEM (standard error of mean). Statistical significance was determined by two-way ANOVA with multiple comparisons. P -values from statistical comparisons between indicated groups are shown above the bars.
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    A Schematic illustration showing how PC/PE imbalance due to impaired SAM-dependent methylation leads to ER stress and mitochondrial dysfunction. The illustration was created in BioRender. Zang, Y. (2026) https://BioRender.com/juq60sz . B Heatmap of individual PC/PE species ratios from hepatic lipidomic analysis comparing sham-GAN + CCl 4 and VSG-GAN + CCl 4 groups ( n = 9, 10, respectively). C Total hepatic PC/PE ratio in sham-GAN + CCl 4 , and VSG-GAN+CCl 4 groups ( n = 9, 10 respectively). D Western blot analysis of ER stress markers (P-PERK, PERK, <t>P-eIF2α,</t> eIF2α) in liver samples from sham-Chow, sham-GAN+CCl 4 and VSG-GAN + CCl 4 groups ( n = 3 per group). E Densitometric quantification of p-PERK/PERK and p-eIF2α/eIF2α ratios from panel ( D ). F Quantification of mitochondrial ROS (mROS) production in isolated hepatocytes from indicated groups ( n = 3 per group). G Quantification of hepatic malondialdehyde (MDA) levels as a marker of lipid peroxidation ( n = 6, 8, 8 for sham-Chow, sham-GAN + CCl 4 , and VSG-GAN + CCl 4 , respectively). H Quantification of mitochondrial DNA to nuclear DNA ratio indicating mitochondrial content in liver samples ( n = 6, 8, 8 for sham-Chow, sham-GAN + CCl 4 , and VSG-GAN + CCl 4 , respectively). I Quantification of hepatic ATP content ( n = 6, 8, 8 for sham-Chow, sham-GAN + CCl 4 , and VSG-GAN + CCl 4 , respectively). All data are presented as mean ± SEM (standard error of mean). Statistical significance was determined by two-way ANOVA with multiple comparisons. P -values from statistical comparisons between indicated groups are shown above the bars.
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    A Schematic illustration showing how PC/PE imbalance due to impaired SAM-dependent methylation leads to ER stress and mitochondrial dysfunction. The illustration was created in BioRender. Zang, Y. (2026) https://BioRender.com/juq60sz . B Heatmap of individual PC/PE species ratios from hepatic lipidomic analysis comparing sham-GAN + CCl 4 and VSG-GAN + CCl 4 groups ( n = 9, 10, respectively). C Total hepatic PC/PE ratio in sham-GAN + CCl 4 , and VSG-GAN+CCl 4 groups ( n = 9, 10 respectively). D Western blot analysis of ER stress markers (P-PERK, PERK, <t>P-eIF2α,</t> eIF2α) in liver samples from sham-Chow, sham-GAN+CCl 4 and VSG-GAN + CCl 4 groups ( n = 3 per group). E Densitometric quantification of p-PERK/PERK and p-eIF2α/eIF2α ratios from panel ( D ). F Quantification of mitochondrial ROS (mROS) production in isolated hepatocytes from indicated groups ( n = 3 per group). G Quantification of hepatic malondialdehyde (MDA) levels as a marker of lipid peroxidation ( n = 6, 8, 8 for sham-Chow, sham-GAN + CCl 4 , and VSG-GAN + CCl 4 , respectively). H Quantification of mitochondrial DNA to nuclear DNA ratio indicating mitochondrial content in liver samples ( n = 6, 8, 8 for sham-Chow, sham-GAN + CCl 4 , and VSG-GAN + CCl 4 , respectively). I Quantification of hepatic ATP content ( n = 6, 8, 8 for sham-Chow, sham-GAN + CCl 4 , and VSG-GAN + CCl 4 , respectively). All data are presented as mean ± SEM (standard error of mean). Statistical significance was determined by two-way ANOVA with multiple comparisons. P -values from statistical comparisons between indicated groups are shown above the bars.
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    A Schematic illustration showing how PC/PE imbalance due to impaired SAM-dependent methylation leads to ER stress and mitochondrial dysfunction. The illustration was created in BioRender. Zang, Y. (2026) https://BioRender.com/juq60sz . B Heatmap of individual PC/PE species ratios from hepatic lipidomic analysis comparing sham-GAN + CCl 4 and VSG-GAN + CCl 4 groups ( n = 9, 10, respectively). C Total hepatic PC/PE ratio in sham-GAN + CCl 4 , and VSG-GAN+CCl 4 groups ( n = 9, 10 respectively). D Western blot analysis of ER stress markers (P-PERK, PERK, <t>P-eIF2α,</t> eIF2α) in liver samples from sham-Chow, sham-GAN+CCl 4 and VSG-GAN + CCl 4 groups ( n = 3 per group). E Densitometric quantification of p-PERK/PERK and p-eIF2α/eIF2α ratios from panel ( D ). F Quantification of mitochondrial ROS (mROS) production in isolated hepatocytes from indicated groups ( n = 3 per group). G Quantification of hepatic malondialdehyde (MDA) levels as a marker of lipid peroxidation ( n = 6, 8, 8 for sham-Chow, sham-GAN + CCl 4 , and VSG-GAN + CCl 4 , respectively). H Quantification of mitochondrial DNA to nuclear DNA ratio indicating mitochondrial content in liver samples ( n = 6, 8, 8 for sham-Chow, sham-GAN + CCl 4 , and VSG-GAN + CCl 4 , respectively). I Quantification of hepatic ATP content ( n = 6, 8, 8 for sham-Chow, sham-GAN + CCl 4 , and VSG-GAN + CCl 4 , respectively). All data are presented as mean ± SEM (standard error of mean). Statistical significance was determined by two-way ANOVA with multiple comparisons. P -values from statistical comparisons between indicated groups are shown above the bars.
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    A Schematic illustration showing how PC/PE imbalance due to impaired SAM-dependent methylation leads to ER stress and mitochondrial dysfunction. The illustration was created in BioRender. Zang, Y. (2026) https://BioRender.com/juq60sz . B Heatmap of individual PC/PE species ratios from hepatic lipidomic analysis comparing sham-GAN + CCl 4 and VSG-GAN + CCl 4 groups ( n = 9, 10, respectively). C Total hepatic PC/PE ratio in sham-GAN + CCl 4 , and VSG-GAN+CCl 4 groups ( n = 9, 10 respectively). D Western blot analysis of ER stress markers (P-PERK, PERK, <t>P-eIF2α,</t> eIF2α) in liver samples from sham-Chow, sham-GAN+CCl 4 and VSG-GAN + CCl 4 groups ( n = 3 per group). E Densitometric quantification of p-PERK/PERK and p-eIF2α/eIF2α ratios from panel ( D ). F Quantification of mitochondrial ROS (mROS) production in isolated hepatocytes from indicated groups ( n = 3 per group). G Quantification of hepatic malondialdehyde (MDA) levels as a marker of lipid peroxidation ( n = 6, 8, 8 for sham-Chow, sham-GAN + CCl 4 , and VSG-GAN + CCl 4 , respectively). H Quantification of mitochondrial DNA to nuclear DNA ratio indicating mitochondrial content in liver samples ( n = 6, 8, 8 for sham-Chow, sham-GAN + CCl 4 , and VSG-GAN + CCl 4 , respectively). I Quantification of hepatic ATP content ( n = 6, 8, 8 for sham-Chow, sham-GAN + CCl 4 , and VSG-GAN + CCl 4 , respectively). All data are presented as mean ± SEM (standard error of mean). Statistical significance was determined by two-way ANOVA with multiple comparisons. P -values from statistical comparisons between indicated groups are shown above the bars.
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    SAR405 inhibits autophagy during erythropoiesis (A) RT-qPCR in WT murine splenic erythroblast cells (Lineage − CD71 + ) 3 days after acute hemolytic anemia. N = 3 mice per group. Error bars represent standard error of mean (SEM). ∗∗∗ p < 0.001 (two-tailed unpaired Student’s t test). (B) RT-qPCR in murine splenic erythroblasts (lineage − CD71 + Kit + ) cells isolated from control (WT) or Berkeley Sickle Cell Disease (SCD) mice ( N = 3 per genotype). Error bars represent standard error of mean (SEM). ∗ p < 0.05; ∗∗ p < 0.01 (two-tailed unpaired Student’s t test). Target mRNA expression levels were normalized to 18S rRNA. (C) Western blotting of Lineage − CD71 + cells isolated from control (PBS-treated) and anemia (PHZ-treated) spleens for autophagy-related proteins. N = 3 WT mice per experimental group. (D) Semi-quantitative densitometry of p -Ampk (Thr172) relative to total Ampk and p <t>-eIF2α</t> <t>(Ser51)</t> relative to total eIF2α ( N = 3 mice). Error bars represent standard error of mean (SEM). ∗ p < 0.05; ∗∗∗ p < 0.001 (two-tailed unpaired Student’s t test). (E) Experimental layout of splenic EPCs isolated from Samd14 fl/fl (WT) and Samd14 fl/fl;Vav−Cre (S14-CKO) mice post PHZ-induced anemia. EPCs were retrovirally infected to express the GFP-LC3-RFP probe (Addgene #84573). After 24 h, cells were treated with VPS34 inhibitor SAR405 for an additional 48 h before analysis by flow cytometry. (F) Schematic representation of autophagic flux measurement using GFP-LC3-RFP probe. (G) Gating strategy for GFP and RFP signal measurement by flow cytometry. Progressively maturing erythroid cells (R1-R4) were gated within a viable RFP-expressing cell population using cell surface markers CD71 and TER119. (H) Quantitation of GFP/RFP median fluorescence intensity ratio calculated within R1 (CD71 low/med TER119 - ), R2 (CD71 high TER119 - ), R3 (CD71 high TER119 + ), and R4 (CD71 low/neg TER119 + ) populations in DMSO or SAR405-treated WT cultures ( n = 3 mice). Error bars represent standard error of mean (SEM). ∗∗∗ p < 0.001 (One-way ANOVA). (I) Quantitation of GFP/RFP median fluorescence intensity ratio calculated within R2 (CD71 high TER119 - ) and R4 (CD71 low/neg TER119 + ) populations in DMSO or SAR405-treated WT and S14-CKO cultures ( n = 3 mice per group). Error bars represent standard deviation (SD). ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001 (two-way ANOVA).
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    Image Search Results


    Effect of in-feed supplementation of SP on eIF2α in breast of heat-stressed broilers. The levels of eIF2α protein (A, B) and mRNA (C) were determined by Western blot and qPCR, respectively. Data are presented as mean ± SD (n=6 birds/group) and analyzed by Two-way ANOVA and Tukey’s HSD multiple comparison test. Different superscript letters indicate significant difference at P < 0.05. The immunoblot is a representative illustration of 6 biological replicates. eIF2α, eukaryotic translation initiation factor 2 alpha subunit; HS, heat stress; TN, thermoneutral; PF, pair fed.

    Journal: Frontiers in Physiology

    Article Title: In-feed Arthrospira (Spirulina) platensis supplementation enhances breast yield in heat-stressed broilers through modulation of proteostasis network

    doi: 10.3389/fphys.2026.1912121

    Figure Lengend Snippet: Effect of in-feed supplementation of SP on eIF2α in breast of heat-stressed broilers. The levels of eIF2α protein (A, B) and mRNA (C) were determined by Western blot and qPCR, respectively. Data are presented as mean ± SD (n=6 birds/group) and analyzed by Two-way ANOVA and Tukey’s HSD multiple comparison test. Different superscript letters indicate significant difference at P < 0.05. The immunoblot is a representative illustration of 6 biological replicates. eIF2α, eukaryotic translation initiation factor 2 alpha subunit; HS, heat stress; TN, thermoneutral; PF, pair fed.

    Article Snippet: Primary antibodies used were: rabbit anti-mechanistic target of rapamycin (mTOR, #2972, Cell Signaling Technology, Danvers, MA), rabbit anti-phospho-mTOR Ser2481 (#AP0978, ABclonal, Woburn, MA), rabbit anti-phospho-mTOR Ser2448 (#AP0115, ABclonal, Woburn, MA), rabbit anti-p70 S6 kinase α (S6Kα, #sc-230, Santa Cruz Biotechnology, Dallas, TX), rabbit anti-phospho-p70 S6 kinase α Thr 421/Ser 424 (p-S6Kα, #sc-7984-R, Santa Cruz Biotechnology, Dallas, TX), rabbit anti-phospho S6 ribosomal protein Ser235/236 (p-S6, #AP0538, ABclonal, Woburn, MA), rabbit anti-phospho eukaryotic translation initiation factor 2 alpha Ser51 (p-eIF2α, #AP0692, ABclonal, Woburn, MA), rabbit anti-eIF2α (#A0764, ABclonal, Woburn, MA), rabbit anti-ubiquitin (#A19686, ABclonal, Woburn, MA), mouse anti-heat shock protein 70 (HSP70, #MA3-007, Thermo Scientific, Waltham, MA), mouse anti-HSP90 (#ab59459, Abcam, Waltham, MA), and rabbit anti-HSP60 (#ARP61107_P050, Aviva Systems Biology San Diego, CA).

    Techniques: Western Blot, Comparison

    Effect of BA and QCT treatments on eIF2α pathway in CPEM. The levels of eIF2α protein (A-D) and mRNA (E-G) were determined by Western blot and qPCR, respectively. Data are presented as mean ± SD (n=6 replicates/treatment) and analyzed by Two-way ANOVA and Tukey’s HSD multiple comparison test. *, **, *** indicate significant differences at P < 0.05; P < 0.01; and P < 0.001, respectively. The immunoblot is a representative illustration of 6 biological replicates. C, control; BA, benzoic acid: eIF2α, eukaryotic translation initiation factor 2 alpha subunit; QCT, quercetin.

    Journal: Frontiers in Physiology

    Article Title: In-feed Arthrospira (Spirulina) platensis supplementation enhances breast yield in heat-stressed broilers through modulation of proteostasis network

    doi: 10.3389/fphys.2026.1912121

    Figure Lengend Snippet: Effect of BA and QCT treatments on eIF2α pathway in CPEM. The levels of eIF2α protein (A-D) and mRNA (E-G) were determined by Western blot and qPCR, respectively. Data are presented as mean ± SD (n=6 replicates/treatment) and analyzed by Two-way ANOVA and Tukey’s HSD multiple comparison test. *, **, *** indicate significant differences at P < 0.05; P < 0.01; and P < 0.001, respectively. The immunoblot is a representative illustration of 6 biological replicates. C, control; BA, benzoic acid: eIF2α, eukaryotic translation initiation factor 2 alpha subunit; QCT, quercetin.

    Article Snippet: Primary antibodies used were: rabbit anti-mechanistic target of rapamycin (mTOR, #2972, Cell Signaling Technology, Danvers, MA), rabbit anti-phospho-mTOR Ser2481 (#AP0978, ABclonal, Woburn, MA), rabbit anti-phospho-mTOR Ser2448 (#AP0115, ABclonal, Woburn, MA), rabbit anti-p70 S6 kinase α (S6Kα, #sc-230, Santa Cruz Biotechnology, Dallas, TX), rabbit anti-phospho-p70 S6 kinase α Thr 421/Ser 424 (p-S6Kα, #sc-7984-R, Santa Cruz Biotechnology, Dallas, TX), rabbit anti-phospho S6 ribosomal protein Ser235/236 (p-S6, #AP0538, ABclonal, Woburn, MA), rabbit anti-phospho eukaryotic translation initiation factor 2 alpha Ser51 (p-eIF2α, #AP0692, ABclonal, Woburn, MA), rabbit anti-eIF2α (#A0764, ABclonal, Woburn, MA), rabbit anti-ubiquitin (#A19686, ABclonal, Woburn, MA), mouse anti-heat shock protein 70 (HSP70, #MA3-007, Thermo Scientific, Waltham, MA), mouse anti-HSP90 (#ab59459, Abcam, Waltham, MA), and rabbit anti-HSP60 (#ARP61107_P050, Aviva Systems Biology San Diego, CA).

    Techniques: Western Blot, Comparison, Control

    Sources of primary antibodies, dilutions for Western blotting and immunolocalization. NA, not applicable.

    Journal: Cells

    Article Title: Ivermectin Inhibits Stress Granule Clearance by Blocking the De Novo Synthesis of Hsp70 in Neuroblastoma Cells

    doi: 10.3390/cells15171623

    Figure Lengend Snippet: Sources of primary antibodies, dilutions for Western blotting and immunolocalization. NA, not applicable.

    Article Snippet: p-eIF2α (S51) , ABclonal, Woburn, MA, USA , AP0745 , 1:1000 , NA.

    Techniques: Western Blot

    A Schematic illustration showing how PC/PE imbalance due to impaired SAM-dependent methylation leads to ER stress and mitochondrial dysfunction. The illustration was created in BioRender. Zang, Y. (2026) https://BioRender.com/juq60sz . B Heatmap of individual PC/PE species ratios from hepatic lipidomic analysis comparing sham-GAN + CCl 4 and VSG-GAN + CCl 4 groups ( n = 9, 10, respectively). C Total hepatic PC/PE ratio in sham-GAN + CCl 4 , and VSG-GAN+CCl 4 groups ( n = 9, 10 respectively). D Western blot analysis of ER stress markers (P-PERK, PERK, P-eIF2α, eIF2α) in liver samples from sham-Chow, sham-GAN+CCl 4 and VSG-GAN + CCl 4 groups ( n = 3 per group). E Densitometric quantification of p-PERK/PERK and p-eIF2α/eIF2α ratios from panel ( D ). F Quantification of mitochondrial ROS (mROS) production in isolated hepatocytes from indicated groups ( n = 3 per group). G Quantification of hepatic malondialdehyde (MDA) levels as a marker of lipid peroxidation ( n = 6, 8, 8 for sham-Chow, sham-GAN + CCl 4 , and VSG-GAN + CCl 4 , respectively). H Quantification of mitochondrial DNA to nuclear DNA ratio indicating mitochondrial content in liver samples ( n = 6, 8, 8 for sham-Chow, sham-GAN + CCl 4 , and VSG-GAN + CCl 4 , respectively). I Quantification of hepatic ATP content ( n = 6, 8, 8 for sham-Chow, sham-GAN + CCl 4 , and VSG-GAN + CCl 4 , respectively). All data are presented as mean ± SEM (standard error of mean). Statistical significance was determined by two-way ANOVA with multiple comparisons. P -values from statistical comparisons between indicated groups are shown above the bars.

    Journal: Nature Communications

    Article Title: Bariatric surgery resolves MASH by enhancing MAT1A-dependent one-carbon metabolism

    doi: 10.1038/s41467-026-75822-y

    Figure Lengend Snippet: A Schematic illustration showing how PC/PE imbalance due to impaired SAM-dependent methylation leads to ER stress and mitochondrial dysfunction. The illustration was created in BioRender. Zang, Y. (2026) https://BioRender.com/juq60sz . B Heatmap of individual PC/PE species ratios from hepatic lipidomic analysis comparing sham-GAN + CCl 4 and VSG-GAN + CCl 4 groups ( n = 9, 10, respectively). C Total hepatic PC/PE ratio in sham-GAN + CCl 4 , and VSG-GAN+CCl 4 groups ( n = 9, 10 respectively). D Western blot analysis of ER stress markers (P-PERK, PERK, P-eIF2α, eIF2α) in liver samples from sham-Chow, sham-GAN+CCl 4 and VSG-GAN + CCl 4 groups ( n = 3 per group). E Densitometric quantification of p-PERK/PERK and p-eIF2α/eIF2α ratios from panel ( D ). F Quantification of mitochondrial ROS (mROS) production in isolated hepatocytes from indicated groups ( n = 3 per group). G Quantification of hepatic malondialdehyde (MDA) levels as a marker of lipid peroxidation ( n = 6, 8, 8 for sham-Chow, sham-GAN + CCl 4 , and VSG-GAN + CCl 4 , respectively). H Quantification of mitochondrial DNA to nuclear DNA ratio indicating mitochondrial content in liver samples ( n = 6, 8, 8 for sham-Chow, sham-GAN + CCl 4 , and VSG-GAN + CCl 4 , respectively). I Quantification of hepatic ATP content ( n = 6, 8, 8 for sham-Chow, sham-GAN + CCl 4 , and VSG-GAN + CCl 4 , respectively). All data are presented as mean ± SEM (standard error of mean). Statistical significance was determined by two-way ANOVA with multiple comparisons. P -values from statistical comparisons between indicated groups are shown above the bars.

    Article Snippet: ABclonal: P-eIF2α (1:1000, AP0692), and eIF2α (1:1000, A21221).

    Techniques: Methylation, Western Blot, Isolation, Marker

    SAR405 inhibits autophagy during erythropoiesis (A) RT-qPCR in WT murine splenic erythroblast cells (Lineage − CD71 + ) 3 days after acute hemolytic anemia. N = 3 mice per group. Error bars represent standard error of mean (SEM). ∗∗∗ p < 0.001 (two-tailed unpaired Student’s t test). (B) RT-qPCR in murine splenic erythroblasts (lineage − CD71 + Kit + ) cells isolated from control (WT) or Berkeley Sickle Cell Disease (SCD) mice ( N = 3 per genotype). Error bars represent standard error of mean (SEM). ∗ p < 0.05; ∗∗ p < 0.01 (two-tailed unpaired Student’s t test). Target mRNA expression levels were normalized to 18S rRNA. (C) Western blotting of Lineage − CD71 + cells isolated from control (PBS-treated) and anemia (PHZ-treated) spleens for autophagy-related proteins. N = 3 WT mice per experimental group. (D) Semi-quantitative densitometry of p -Ampk (Thr172) relative to total Ampk and p -eIF2α (Ser51) relative to total eIF2α ( N = 3 mice). Error bars represent standard error of mean (SEM). ∗ p < 0.05; ∗∗∗ p < 0.001 (two-tailed unpaired Student’s t test). (E) Experimental layout of splenic EPCs isolated from Samd14 fl/fl (WT) and Samd14 fl/fl;Vav−Cre (S14-CKO) mice post PHZ-induced anemia. EPCs were retrovirally infected to express the GFP-LC3-RFP probe (Addgene #84573). After 24 h, cells were treated with VPS34 inhibitor SAR405 for an additional 48 h before analysis by flow cytometry. (F) Schematic representation of autophagic flux measurement using GFP-LC3-RFP probe. (G) Gating strategy for GFP and RFP signal measurement by flow cytometry. Progressively maturing erythroid cells (R1-R4) were gated within a viable RFP-expressing cell population using cell surface markers CD71 and TER119. (H) Quantitation of GFP/RFP median fluorescence intensity ratio calculated within R1 (CD71 low/med TER119 - ), R2 (CD71 high TER119 - ), R3 (CD71 high TER119 + ), and R4 (CD71 low/neg TER119 + ) populations in DMSO or SAR405-treated WT cultures ( n = 3 mice). Error bars represent standard error of mean (SEM). ∗∗∗ p < 0.001 (One-way ANOVA). (I) Quantitation of GFP/RFP median fluorescence intensity ratio calculated within R2 (CD71 high TER119 - ) and R4 (CD71 low/neg TER119 + ) populations in DMSO or SAR405-treated WT and S14-CKO cultures ( n = 3 mice per group). Error bars represent standard deviation (SD). ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001 (two-way ANOVA).

    Journal: iScience

    Article Title: A stress-induced PI3P complex controls autophagy in erythroid precursors

    doi: 10.1016/j.isci.2026.115182

    Figure Lengend Snippet: SAR405 inhibits autophagy during erythropoiesis (A) RT-qPCR in WT murine splenic erythroblast cells (Lineage − CD71 + ) 3 days after acute hemolytic anemia. N = 3 mice per group. Error bars represent standard error of mean (SEM). ∗∗∗ p < 0.001 (two-tailed unpaired Student’s t test). (B) RT-qPCR in murine splenic erythroblasts (lineage − CD71 + Kit + ) cells isolated from control (WT) or Berkeley Sickle Cell Disease (SCD) mice ( N = 3 per genotype). Error bars represent standard error of mean (SEM). ∗ p < 0.05; ∗∗ p < 0.01 (two-tailed unpaired Student’s t test). Target mRNA expression levels were normalized to 18S rRNA. (C) Western blotting of Lineage − CD71 + cells isolated from control (PBS-treated) and anemia (PHZ-treated) spleens for autophagy-related proteins. N = 3 WT mice per experimental group. (D) Semi-quantitative densitometry of p -Ampk (Thr172) relative to total Ampk and p -eIF2α (Ser51) relative to total eIF2α ( N = 3 mice). Error bars represent standard error of mean (SEM). ∗ p < 0.05; ∗∗∗ p < 0.001 (two-tailed unpaired Student’s t test). (E) Experimental layout of splenic EPCs isolated from Samd14 fl/fl (WT) and Samd14 fl/fl;Vav−Cre (S14-CKO) mice post PHZ-induced anemia. EPCs were retrovirally infected to express the GFP-LC3-RFP probe (Addgene #84573). After 24 h, cells were treated with VPS34 inhibitor SAR405 for an additional 48 h before analysis by flow cytometry. (F) Schematic representation of autophagic flux measurement using GFP-LC3-RFP probe. (G) Gating strategy for GFP and RFP signal measurement by flow cytometry. Progressively maturing erythroid cells (R1-R4) were gated within a viable RFP-expressing cell population using cell surface markers CD71 and TER119. (H) Quantitation of GFP/RFP median fluorescence intensity ratio calculated within R1 (CD71 low/med TER119 - ), R2 (CD71 high TER119 - ), R3 (CD71 high TER119 + ), and R4 (CD71 low/neg TER119 + ) populations in DMSO or SAR405-treated WT cultures ( n = 3 mice). Error bars represent standard error of mean (SEM). ∗∗∗ p < 0.001 (One-way ANOVA). (I) Quantitation of GFP/RFP median fluorescence intensity ratio calculated within R2 (CD71 high TER119 - ) and R4 (CD71 low/neg TER119 + ) populations in DMSO or SAR405-treated WT and S14-CKO cultures ( n = 3 mice per group). Error bars represent standard deviation (SD). ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001 (two-way ANOVA).

    Article Snippet: Anti-phospho-eIF2α (Ser51) (clone 119A11, rabbit monoclonal) , Cell Signaling Technology , Cat.: #3597; RRID: AB_390740.

    Techniques: Quantitative RT-PCR, Two Tailed Test, Isolation, Control, Expressing, Western Blot, Infection, Flow Cytometry, Quantitation Assay, Fluorescence, Standard Deviation