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gene exp atf4 hs00909569 g1  (Thermo Fisher)


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    Structured Review

    Thermo Fisher gene exp atf4 hs00909569 g1
    RNA sequencing reveals that tomatidine modulates <t>ATF4-dependent</t> ER stress genes in pancreatic cancer cells Human and murine PDAC cell lines were treated for 40 h with 6.4 μg/mL tomatidine, RNA was isolated, and RNA sequencing was performed to analyze differences in gene regulation. N = 3 biological separate experiments. (A–D) (A) Heatmap of all genes and how they change in tomatidine-treated vs. untreated cells for Panc1 cells (FDR<0.05). Volcano plot highlighting ER stress-related genes in (B) Panc1 and (C) MT5 cells. (Upregulated genes on the right of the central axis and vice versa with higher fold change as we go away from the origin on y axis) (D) IPA upstream analysis of ATF4-related genes in Panc1 cells. (E) Heatmap elucidating targeting of ATF4-related genes in treated vs. untreated Panc1 cells. (Fold change: +2.5 to −1.5; FDR<0.03). (F) Top 10 pathways focusing on UPR, ATF4, and ER stress via Reactome analysis of the RNA-sequencing data of treated vs. untreated samples for Panc1 cells. (FDR< 0.03).
    Gene Exp Atf4 Hs00909569 G1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/transcriptome+sequencing+experiment/Gene+Exp%2E+ATF4%2C+Hs00909569_g1/pmc10405072-368-30-18
    Average 94 stars, based on 1 article reviews
    gene exp atf4 hs00909569 g1 - by Bioz Stars, 2026-09
    94/100 stars

    Images

    1) Product Images from "Tomatidine targets ATF4-dependent signaling and induces ferroptosis to limit pancreatic cancer progression"

    Article Title: Tomatidine targets ATF4-dependent signaling and induces ferroptosis to limit pancreatic cancer progression

    Journal: iScience

    doi: 10.1016/j.isci.2023.107408

    RNA sequencing reveals that tomatidine modulates ATF4-dependent ER stress genes in pancreatic cancer cells Human and murine PDAC cell lines were treated for 40 h with 6.4 μg/mL tomatidine, RNA was isolated, and RNA sequencing was performed to analyze differences in gene regulation. N = 3 biological separate experiments. (A–D) (A) Heatmap of all genes and how they change in tomatidine-treated vs. untreated cells for Panc1 cells (FDR<0.05). Volcano plot highlighting ER stress-related genes in (B) Panc1 and (C) MT5 cells. (Upregulated genes on the right of the central axis and vice versa with higher fold change as we go away from the origin on y axis) (D) IPA upstream analysis of ATF4-related genes in Panc1 cells. (E) Heatmap elucidating targeting of ATF4-related genes in treated vs. untreated Panc1 cells. (Fold change: +2.5 to −1.5; FDR<0.03). (F) Top 10 pathways focusing on UPR, ATF4, and ER stress via Reactome analysis of the RNA-sequencing data of treated vs. untreated samples for Panc1 cells. (FDR< 0.03).
    Figure Legend Snippet: RNA sequencing reveals that tomatidine modulates ATF4-dependent ER stress genes in pancreatic cancer cells Human and murine PDAC cell lines were treated for 40 h with 6.4 μg/mL tomatidine, RNA was isolated, and RNA sequencing was performed to analyze differences in gene regulation. N = 3 biological separate experiments. (A–D) (A) Heatmap of all genes and how they change in tomatidine-treated vs. untreated cells for Panc1 cells (FDR<0.05). Volcano plot highlighting ER stress-related genes in (B) Panc1 and (C) MT5 cells. (Upregulated genes on the right of the central axis and vice versa with higher fold change as we go away from the origin on y axis) (D) IPA upstream analysis of ATF4-related genes in Panc1 cells. (E) Heatmap elucidating targeting of ATF4-related genes in treated vs. untreated Panc1 cells. (Fold change: +2.5 to −1.5; FDR<0.03). (F) Top 10 pathways focusing on UPR, ATF4, and ER stress via Reactome analysis of the RNA-sequencing data of treated vs. untreated samples for Panc1 cells. (FDR< 0.03).

    Techniques Used: RNA Sequencing, Isolation

    ATF4 expression in PDAC (A) KM plotter was utilized to examine ATF4 expression in human PDAC specimens examining high vs. low expression compared to overall survival. (B) Data distribution elucidating significance of the overall survival curve prepared by KM Plotter. (C and D) (C) scRNA-seq datasets of pancreatic cancer tissue from metastatic patients were obtained from NIH dbGAP (accession phs002045.v1.p1) and (D) analyzed for ATF4 expression. (See also <xref ref-type=Table S1 ). (E) Pancreatic tumor tissue from KPC mice were stained by multiplex IF and imaged using Akoya Vectra Polaris and the Phenochart software to analyze ATF4 in multiple cellular compartments. Markers used to determine ATF4 (white), epithelial cells (CK19; red), stroma (αSMA; orange and PDGFRβ; Green), CD8 T+ cells (yellow), and macrophages (F4/80; turquoise). Scale bar = 100 μm. " title="ATF4 expression in PDAC (A) KM plotter was utilized ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: ATF4 expression in PDAC (A) KM plotter was utilized to examine ATF4 expression in human PDAC specimens examining high vs. low expression compared to overall survival. (B) Data distribution elucidating significance of the overall survival curve prepared by KM Plotter. (C and D) (C) scRNA-seq datasets of pancreatic cancer tissue from metastatic patients were obtained from NIH dbGAP (accession phs002045.v1.p1) and (D) analyzed for ATF4 expression. (See also Table S1 ). (E) Pancreatic tumor tissue from KPC mice were stained by multiplex IF and imaged using Akoya Vectra Polaris and the Phenochart software to analyze ATF4 in multiple cellular compartments. Markers used to determine ATF4 (white), epithelial cells (CK19; red), stroma (αSMA; orange and PDGFRβ; Green), CD8 T+ cells (yellow), and macrophages (F4/80; turquoise). Scale bar = 100 μm.

    Techniques Used: Expressing, Staining, Multiplex Assay, Software

    Tomatidine inhibits ATF4-dependent signaling in PDAC (A and B) (A) MiaPaca-2 tumor cells were treated with tomatidine for 72 h and cell lysates were immunoblotted for ATF4, 4EBP1, and phospho-4EBP1(p-4EBP1) protein expression and (B) p-4EBP1/4EBP1 levels quantified by densitometry. (C) Immunofluorescence (IF) was performed on vehicle (DMSO) or tomatidine-treated Panc1 cells to track ATF4 (FITC-Green) translocation from nucleus (DAPI-Blue) to cytoplasm. Scale bar = 50 μm. (D–G) (D) Nuclear to cytoplasmic translocation was quantified. Panc1 cells were treated with vehicle (DMSO) or tomatidine for 40 h and ATF4 transcriptional activity was analyzed by chromatin immunoprecipitation (ChIP) qPCR evaluating binding of ATF4 to the downstream promoter regions of (E) eIF4EBP1 (F) CHOP- B site and (G) ASNS. Data are reported as the means + SEMs. n = 3 or more independent biological replicates (4B, One-way ANOVA with Tukey’s test for pairwise comparisons was used to analyze the data; 4D-G, Two-tailed independent student’s test was used to analyze the data, ∗p < 0.05).
    Figure Legend Snippet: Tomatidine inhibits ATF4-dependent signaling in PDAC (A and B) (A) MiaPaca-2 tumor cells were treated with tomatidine for 72 h and cell lysates were immunoblotted for ATF4, 4EBP1, and phospho-4EBP1(p-4EBP1) protein expression and (B) p-4EBP1/4EBP1 levels quantified by densitometry. (C) Immunofluorescence (IF) was performed on vehicle (DMSO) or tomatidine-treated Panc1 cells to track ATF4 (FITC-Green) translocation from nucleus (DAPI-Blue) to cytoplasm. Scale bar = 50 μm. (D–G) (D) Nuclear to cytoplasmic translocation was quantified. Panc1 cells were treated with vehicle (DMSO) or tomatidine for 40 h and ATF4 transcriptional activity was analyzed by chromatin immunoprecipitation (ChIP) qPCR evaluating binding of ATF4 to the downstream promoter regions of (E) eIF4EBP1 (F) CHOP- B site and (G) ASNS. Data are reported as the means + SEMs. n = 3 or more independent biological replicates (4B, One-way ANOVA with Tukey’s test for pairwise comparisons was used to analyze the data; 4D-G, Two-tailed independent student’s test was used to analyze the data, ∗p < 0.05).

    Techniques Used: Expressing, Immunofluorescence, Translocation Assay, Activity Assay, Chromatin Immunoprecipitation, ChIP-qPCR, Binding Assay, Two Tailed Test

    In vivo tomatidine treatment inhibits pancreatic tumor growth (A–C) (A) MT5 tumor-bearing C57BL/6 mice (5 mice/group) were treated with 5 mg/kg daily i.p. injections of tomatidine or vehicle control (40% HPBCD) and monitored for tumor growth. (n = 5/group). RNA isolated from the tumor tissues of MT5 tumor-bearing C57BL/6 mice treated with vehicle or 5 mg/kg daily i.p. injections of tomatidine were assessed for (B) ATF4 and (C) eIF4EBP1 expression via qPCR. Data are reported as the means + SEMs. n = 5 mice per group. (5A, mixed between-within subjects ANOVA shows a significant interaction between days of treatment and group; 5B-C, Two-tailed independent student’s test was used to analyze the data, ∗p < 0.05).
    Figure Legend Snippet: In vivo tomatidine treatment inhibits pancreatic tumor growth (A–C) (A) MT5 tumor-bearing C57BL/6 mice (5 mice/group) were treated with 5 mg/kg daily i.p. injections of tomatidine or vehicle control (40% HPBCD) and monitored for tumor growth. (n = 5/group). RNA isolated from the tumor tissues of MT5 tumor-bearing C57BL/6 mice treated with vehicle or 5 mg/kg daily i.p. injections of tomatidine were assessed for (B) ATF4 and (C) eIF4EBP1 expression via qPCR. Data are reported as the means + SEMs. n = 5 mice per group. (5A, mixed between-within subjects ANOVA shows a significant interaction between days of treatment and group; 5B-C, Two-tailed independent student’s test was used to analyze the data, ∗p < 0.05).

    Techniques Used: In Vivo, Control, Isolation, Expressing, Two Tailed Test

    Tomatidine mediated inhibition of ATF4 signaling can increase sensitivity to ferroptotic cell death in PDAC (A) Panc-1 tumor cells were treated with tomatidine (6.4 μg/mL) and analyzed by RNA sequencing. Ingenuity pathway analysis of the regulated genes suggested ferroptosis as a top hit for tomatidine-treated cells. (B–D) (B) Pancreatic cancer cells were treated with vehicle (DMSO), erastin (to induce ferroptosis), ferrostatin-1 (to inhibit ferroptosis), tomatidine, or in different combinations and lipid peroxidation of (C) Panc-1 and (D) MiaPaca-2 was analyzed by flow cytometry using Bodipy-11. (E) Panc-1 and MiaPaca-2 cells treated with vehicle (DMSO) or tomatidine and lysates collected after 24 h were immunoblot for GPX4 expression. (F) Panc1 cells were plated overnight and treated with tomatidine for 6 h and then assayed by Seahorse assay to analyze mitochondrial fitness. (G) Schematic showing how tomatidine can regulate ATF4-dependent signaling to induce ferroptosis in pancreatic cancer. Data are reported as the means + SEMs. n = 3 or more independent biological replicates. (7C-D, Two-tailed independent student’s test was used to analyze the data; 7F, Two-tailed independent student’s test was used to analyze the data, ∗p < 0.05, ∗∗p < 0.007).
    Figure Legend Snippet: Tomatidine mediated inhibition of ATF4 signaling can increase sensitivity to ferroptotic cell death in PDAC (A) Panc-1 tumor cells were treated with tomatidine (6.4 μg/mL) and analyzed by RNA sequencing. Ingenuity pathway analysis of the regulated genes suggested ferroptosis as a top hit for tomatidine-treated cells. (B–D) (B) Pancreatic cancer cells were treated with vehicle (DMSO), erastin (to induce ferroptosis), ferrostatin-1 (to inhibit ferroptosis), tomatidine, or in different combinations and lipid peroxidation of (C) Panc-1 and (D) MiaPaca-2 was analyzed by flow cytometry using Bodipy-11. (E) Panc-1 and MiaPaca-2 cells treated with vehicle (DMSO) or tomatidine and lysates collected after 24 h were immunoblot for GPX4 expression. (F) Panc1 cells were plated overnight and treated with tomatidine for 6 h and then assayed by Seahorse assay to analyze mitochondrial fitness. (G) Schematic showing how tomatidine can regulate ATF4-dependent signaling to induce ferroptosis in pancreatic cancer. Data are reported as the means + SEMs. n = 3 or more independent biological replicates. (7C-D, Two-tailed independent student’s test was used to analyze the data; 7F, Two-tailed independent student’s test was used to analyze the data, ∗p < 0.05, ∗∗p < 0.007).

    Techniques Used: Inhibition, RNA Sequencing, Flow Cytometry, Western Blot, Expressing, Two Tailed Test


    Figure Legend Snippet:

    Techniques Used: Control, Recombinant, Viability Assay, Luciferase, ChIP-qPCR, Gene Expression, Software

    Related Articles

    Gene Expression:

    Article Title: Modelling ferroptosis in a human microglial line by sequential exposure to iron and GPX4 inhibition
    Article Snippet: TaqMan Gene Expression Assays (Applied Biosystems, 4331182) and TaqMan Fast Advanced Mastermix (Applied Biosystems, 4444558) were used on a QuantStudio 7 Flex system (ThermoFisher Scientific). .. The following TaqMan assays were used for human gene expression analysis: ATF4 (Hs00909569_g1), ATF6 (Hs00232586_m1), XBP1 (Hs00231936_m1), GPX4 (Hs00989766_g1). .. Relative gene expression was determined via the Comparative CT method normalised to at least 3 housekeeping genes (ACTB (Hs01060665_g1), B2M (Hs00187842_m1), GAPDH (HS02786624_g1), TBP (Hs00427620_m1)).

    Article Title: Apoptosis and motor deficits in SPG76 hereditary spastic paraplegia: Calpain 2 inhibition as therapeutic strategy.
    Article Snippet: RNA was prepared from fibroblast cells using Trizol (Invitrogen, Thermo Fisher Scientific) and 1 μg/samples was reverse-transcribed into cDNA using the Superscript III First Strand Synthesis System for RT-PCR kit (Invitrogen, Thermo Fisher Scientific) and random hexamers. .. The expression levels of GRP78 (HSPA5, Hs99999174_m1), GRP94 (HSP90B1, Hs00427665_g1), ATF4 (Hs00909569_g1), XBP1s (XBP1s, Hs03929085_g1), XBP1 unspliced (XBP1u, Hs00231936_m1), IRE1 (ERN1, Hs00980095_m1) and CHOP (DDIT3, Hs99999172_m1) were analysed by quantitative Real Time PCR on a QuantStudioTM 3 RealTime PCR System (Applied Biosystems, Thermo Fisher Scientific) by using specific TaqManTM Gene Expression Assay and TaqmanTM Universal PCR Master Mix (4324018, Thermo Fisher Scientific). .. GAPDH (Hs02758991_g1) was used as reference gene for normalization.

    other:

    Article Title:
    Article Snippet: TaqMan primer probes used for analysis of UPR and cell death genes: 18s, Hs99999901_s1; CHOP/DDIT3, Hs01090850_m1; GADD34, Hs00169585_m1; BIP, Hs99999174_m1; ATF3, HsYYY; ATF4, Hs00909569_g1; ATF5, HsYYY; NOXA, Hs00560402_m1; PUMA, Hs00248075_m1; BIM, Hs00197982_m1; NBK, Hs00154189_m1, BCL2, HsYYY; TRB3, HsYYY; DR5, HsYYY; ERO1, HsYYY.

    Article Title: Single-cell mRNA-regulation analysis reveals cell type-specific mechanisms of type 2 diabetes
    Article Snippet: TaqMans for human islet were: ATF4 (Hs00909569_g1), ATF6 (Hs00232586_m1), CEBPG (Hs01922818_s1), DDIT3 (Hs99999172_m1), HPRT1 (Hs4326321_m1), INS (Hs02741908_m1), TBP (Hs00427620_m1), and XBP1 (Hs00231936_m1).

    Expressing:

    Article Title: Apoptosis and motor deficits in SPG76 hereditary spastic paraplegia: Calpain 2 inhibition as therapeutic strategy.
    Article Snippet: RNA was prepared from fibroblast cells using Trizol (Invitrogen, Thermo Fisher Scientific) and 1 μg/samples was reverse-transcribed into cDNA using the Superscript III First Strand Synthesis System for RT-PCR kit (Invitrogen, Thermo Fisher Scientific) and random hexamers. .. The expression levels of GRP78 (HSPA5, Hs99999174_m1), GRP94 (HSP90B1, Hs00427665_g1), ATF4 (Hs00909569_g1), XBP1s (XBP1s, Hs03929085_g1), XBP1 unspliced (XBP1u, Hs00231936_m1), IRE1 (ERN1, Hs00980095_m1) and CHOP (DDIT3, Hs99999172_m1) were analysed by quantitative Real Time PCR on a QuantStudioTM 3 RealTime PCR System (Applied Biosystems, Thermo Fisher Scientific) by using specific TaqManTM Gene Expression Assay and TaqmanTM Universal PCR Master Mix (4324018, Thermo Fisher Scientific). .. GAPDH (Hs02758991_g1) was used as reference gene for normalization.

    Article Title: GCN2 and perk kinase inhibitors and methods of use thereof
    Article Snippet: Cells were seeded in 6-well culture plate at 3.0×106 cells per well in 2 mL complete growth medium and incubated with indicated concentrations of Compound 2 at 37° C., 5% CO2, and 95% humidity for 4 h. At the end of the incubation, cells were washed with PBS (Sigma) and RNA was extracted using RNeasy® Plus Kits (Qiagen, Germantown, MD). cDNA was synthesized using the High-Capacity cDNA Reverse Transcription Kit (Applied Biosciences, Beverly Hills, CA), and quantitative PCR was performed on a QuantStudio 3 (Applied Biosciences) using TaqMan Assays (Table 1, Thermo) and TaqMan Fast Advanced Master Mix (Thermo) according to manufacturer's specifications. .. TABLE TaqMan Probes Gene of Interest Assay ID ATF4 Hs00909569_g1 DDIT3 (CHOP) Hs00358796_g1 GPT2 Hs00370287_m1 GUSB (Endogenous Control) Hs00939627_m1 PPP1R15A (GADD34) Hs00169585_m1 VEGFA Hs00900055_m1 Compound 2 unexpectedly induced the expression of UPR/ISR target genes in H929 multiple myeloma cells. ..

    Real-time Polymerase Chain Reaction:

    Article Title: Apoptosis and motor deficits in SPG76 hereditary spastic paraplegia: Calpain 2 inhibition as therapeutic strategy.
    Article Snippet: RNA was prepared from fibroblast cells using Trizol (Invitrogen, Thermo Fisher Scientific) and 1 μg/samples was reverse-transcribed into cDNA using the Superscript III First Strand Synthesis System for RT-PCR kit (Invitrogen, Thermo Fisher Scientific) and random hexamers. .. The expression levels of GRP78 (HSPA5, Hs99999174_m1), GRP94 (HSP90B1, Hs00427665_g1), ATF4 (Hs00909569_g1), XBP1s (XBP1s, Hs03929085_g1), XBP1 unspliced (XBP1u, Hs00231936_m1), IRE1 (ERN1, Hs00980095_m1) and CHOP (DDIT3, Hs99999172_m1) were analysed by quantitative Real Time PCR on a QuantStudioTM 3 RealTime PCR System (Applied Biosystems, Thermo Fisher Scientific) by using specific TaqManTM Gene Expression Assay and TaqmanTM Universal PCR Master Mix (4324018, Thermo Fisher Scientific). .. GAPDH (Hs02758991_g1) was used as reference gene for normalization.

    Polymerase Chain Reaction:

    Article Title: Apoptosis and motor deficits in SPG76 hereditary spastic paraplegia: Calpain 2 inhibition as therapeutic strategy.
    Article Snippet: RNA was prepared from fibroblast cells using Trizol (Invitrogen, Thermo Fisher Scientific) and 1 μg/samples was reverse-transcribed into cDNA using the Superscript III First Strand Synthesis System for RT-PCR kit (Invitrogen, Thermo Fisher Scientific) and random hexamers. .. The expression levels of GRP78 (HSPA5, Hs99999174_m1), GRP94 (HSP90B1, Hs00427665_g1), ATF4 (Hs00909569_g1), XBP1s (XBP1s, Hs03929085_g1), XBP1 unspliced (XBP1u, Hs00231936_m1), IRE1 (ERN1, Hs00980095_m1) and CHOP (DDIT3, Hs99999172_m1) were analysed by quantitative Real Time PCR on a QuantStudioTM 3 RealTime PCR System (Applied Biosystems, Thermo Fisher Scientific) by using specific TaqManTM Gene Expression Assay and TaqmanTM Universal PCR Master Mix (4324018, Thermo Fisher Scientific). .. GAPDH (Hs02758991_g1) was used as reference gene for normalization.

    Control:

    Article Title: GCN2 and perk kinase inhibitors and methods of use thereof
    Article Snippet: Cells were seeded in 6-well culture plate at 3.0×106 cells per well in 2 mL complete growth medium and incubated with indicated concentrations of Compound 2 at 37° C., 5% CO2, and 95% humidity for 4 h. At the end of the incubation, cells were washed with PBS (Sigma) and RNA was extracted using RNeasy® Plus Kits (Qiagen, Germantown, MD). cDNA was synthesized using the High-Capacity cDNA Reverse Transcription Kit (Applied Biosciences, Beverly Hills, CA), and quantitative PCR was performed on a QuantStudio 3 (Applied Biosciences) using TaqMan Assays (Table 1, Thermo) and TaqMan Fast Advanced Master Mix (Thermo) according to manufacturer's specifications. .. TABLE TaqMan Probes Gene of Interest Assay ID ATF4 Hs00909569_g1 DDIT3 (CHOP) Hs00358796_g1 GPT2 Hs00370287_m1 GUSB (Endogenous Control) Hs00939627_m1 PPP1R15A (GADD34) Hs00169585_m1 VEGFA Hs00900055_m1 Compound 2 unexpectedly induced the expression of UPR/ISR target genes in H929 multiple myeloma cells. ..



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