mouse anti atf4  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: Unfolded protein response signaling promotes myeloid cell production and cooperates with oncogenic mutation

doi: 10.1101/2025.09.07.674755

Figure Lengend Snippet: (A) Quantitative RT-PCR analysis of Xbp1 transcripts upon tunicamycin (Tuni) (1.2 μg/ml for 12 hr) or thapsigargin (Thap) (0.05 μM for 6 hr) treatment. Xbp1s: spliced Xbp1 transcript; Xbp1u: unspliced Xbp1 transcript; Ratio: ratio of spliced over unspliced Xbp1 transcript. (B) Representative images and quantification of nuclear ATF4 positive MPP3 cells upon tunicamycin (1.2 μg/ml for 12 hr) or thapsigargin (0.05 μM for 6 hr) treatment. Scale bar: 10 μm. (C) Quantification of myeloid colony formation upon thapsigargin (Thap; 0.05 μM) and IXA4 (10 μM) treatment for 12 hr (n = 4). CFU, colony forming unit; MegE, megakaryocyte/erythrocyte; G (or) M, granulocyte or macrophage; GM; granulocyte/macrophage, GEMM, granulocyte/erythroid/macrophage/megakaryocyte. (D) Quantification of nuclear ATF4 positive MPP3 cells from vehicle versus tunicamycin treated control and Jak2 V617F mice. Data are means ± S.D., and statistical significance was assessed by a two-tailed unpaired Student’s t-test. *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001.

Article Snippet: Cells were then incubated overnight at 4 °C with a rabbit anti-mouse ATF4 (Cell Signaling, 11815S or Abcam, ab216839) primary antibody, washed 3 times with PBS, and incubated for 1 hour at RT with a goat anti-rabbit-A488 (Invitrogen, A32731) secondary antibody.

Techniques: Quantitative RT-PCR, Control, Two Tailed Test

Journal: Cancer immunology, immunotherapy : CII

Article Title: Induction of the activating transcription factor-4 in the intratumoral CD8+ T cells sustains their viability and anti-tumor activities

doi: 10.1007/s00262-022-03286-2

Figure Lengend Snippet: A. qPCR analysis of mRNA levels of Atf4 in mouse CD8+ T cells treated or not with a stress inducer thapsigargin (TG, 100nM, 24 hr) in the presence or absence of CD3/CD28 agonists.

Article Snippet: The following antibodies were used to stain the resulting single cell suspensions: anti-mouse CD8-APC antibody (BioLegend, cat#126614) and anti-human CD8-PE/CY7 antibody (BioLegend, cat#344750), anti-mouse ATF4 primary antibody (Cell signaling Technology, cat#11815s, 1:150), anti-human ATF4 primary antibody (Cell signaling technology, cat#11815s, 1:150), goat anti-rabbit secondary antibody (Invitrogen, cat#A32732, 1:500), anti-CD45-APC/Cy7 (BioLegend, cat#157204), anti-CD3-PE (BioLegend, cat#100206), anti-CD8-AF700 (BioLegend, cat#100730), anti- CD69-BV421 (BioLgend, cat#104545, anti-PD-1-PE/Cy7 (BioLegend, cat#109110) and Annexin V-APC (BioLegend, cat#640941), anti-LAG3-PE (BioLegend, cat#125207), anti-TIGIT-Alexa Fluor 700 (R&D system, cat#FAB72671N), anti-TIM3-BV421 (BioLegend, cat#134019).

Techniques:

Journal: Cancer immunology, immunotherapy : CII

Article Title: Induction of the activating transcription factor-4 in the intratumoral CD8+ T cells sustains their viability and anti-tumor activities

doi: 10.1007/s00262-022-03286-2

Figure Lengend Snippet: A. mRNA levels of Atf4 in mouse CD8+ T cells treated or not with the media conditioned by MC38 tumor cells (TCM) for 48 hr (n=5).

Article Snippet: The following antibodies were used to stain the resulting single cell suspensions: anti-mouse CD8-APC antibody (BioLegend, cat#126614) and anti-human CD8-PE/CY7 antibody (BioLegend, cat#344750), anti-mouse ATF4 primary antibody (Cell signaling Technology, cat#11815s, 1:150), anti-human ATF4 primary antibody (Cell signaling technology, cat#11815s, 1:150), goat anti-rabbit secondary antibody (Invitrogen, cat#A32732, 1:500), anti-CD45-APC/Cy7 (BioLegend, cat#157204), anti-CD3-PE (BioLegend, cat#100206), anti-CD8-AF700 (BioLegend, cat#100730), anti- CD69-BV421 (BioLgend, cat#104545, anti-PD-1-PE/Cy7 (BioLegend, cat#109110) and Annexin V-APC (BioLegend, cat#640941), anti-LAG3-PE (BioLegend, cat#125207), anti-TIGIT-Alexa Fluor 700 (R&D system, cat#FAB72671N), anti-TIM3-BV421 (BioLegend, cat#134019).

Techniques:

Journal: Cancer immunology, immunotherapy : CII

Article Title: Induction of the activating transcription factor-4 in the intratumoral CD8+ T cells sustains their viability and anti-tumor activities

doi: 10.1007/s00262-022-03286-2

Figure Lengend Snippet: A. Frequency of CD3+CD8+ infiltrating T cells in MC38 colon adenocarcinoma tumors from Atf4+/+ or Atf4ΔCD8 mice (n=7).

Article Snippet: The following antibodies were used to stain the resulting single cell suspensions: anti-mouse CD8-APC antibody (BioLegend, cat#126614) and anti-human CD8-PE/CY7 antibody (BioLegend, cat#344750), anti-mouse ATF4 primary antibody (Cell signaling Technology, cat#11815s, 1:150), anti-human ATF4 primary antibody (Cell signaling technology, cat#11815s, 1:150), goat anti-rabbit secondary antibody (Invitrogen, cat#A32732, 1:500), anti-CD45-APC/Cy7 (BioLegend, cat#157204), anti-CD3-PE (BioLegend, cat#100206), anti-CD8-AF700 (BioLegend, cat#100730), anti- CD69-BV421 (BioLgend, cat#104545, anti-PD-1-PE/Cy7 (BioLegend, cat#109110) and Annexin V-APC (BioLegend, cat#640941), anti-LAG3-PE (BioLegend, cat#125207), anti-TIGIT-Alexa Fluor 700 (R&D system, cat#FAB72671N), anti-TIM3-BV421 (BioLegend, cat#134019).

Techniques:

Journal: Cancer immunology, immunotherapy : CII

Article Title: Induction of the activating transcription factor-4 in the intratumoral CD8+ T cells sustains their viability and anti-tumor activities

doi: 10.1007/s00262-022-03286-2

Figure Lengend Snippet: A. Relative ATF4 expression in three CD8+ T cells subtypes isolated from colorectal cancer samples including naïve T cells, effector memory T cells (Tem) and CD103+CD39+ highly active T cells using data from Ref (30). (n=7 for each group).

Article Snippet: The following antibodies were used to stain the resulting single cell suspensions: anti-mouse CD8-APC antibody (BioLegend, cat#126614) and anti-human CD8-PE/CY7 antibody (BioLegend, cat#344750), anti-mouse ATF4 primary antibody (Cell signaling Technology, cat#11815s, 1:150), anti-human ATF4 primary antibody (Cell signaling technology, cat#11815s, 1:150), goat anti-rabbit secondary antibody (Invitrogen, cat#A32732, 1:500), anti-CD45-APC/Cy7 (BioLegend, cat#157204), anti-CD3-PE (BioLegend, cat#100206), anti-CD8-AF700 (BioLegend, cat#100730), anti- CD69-BV421 (BioLgend, cat#104545, anti-PD-1-PE/Cy7 (BioLegend, cat#109110) and Annexin V-APC (BioLegend, cat#640941), anti-LAG3-PE (BioLegend, cat#125207), anti-TIGIT-Alexa Fluor 700 (R&D system, cat#FAB72671N), anti-TIM3-BV421 (BioLegend, cat#134019).

Techniques: Expressing, Isolation

Journal: Cancer immunology, immunotherapy : CII

Article Title: Induction of the activating transcription factor-4 in the intratumoral CD8+ T cells sustains their viability and anti-tumor activities

doi: 10.1007/s00262-022-03286-2

Figure Lengend Snippet: OT-I mice were used in crossings to generate OT-I; Atf4+/+ or OT-I; Atf4ΔCD8 mice.

Article Snippet: The following antibodies were used to stain the resulting single cell suspensions: anti-mouse CD8-APC antibody (BioLegend, cat#126614) and anti-human CD8-PE/CY7 antibody (BioLegend, cat#344750), anti-mouse ATF4 primary antibody (Cell signaling Technology, cat#11815s, 1:150), anti-human ATF4 primary antibody (Cell signaling technology, cat#11815s, 1:150), goat anti-rabbit secondary antibody (Invitrogen, cat#A32732, 1:500), anti-CD45-APC/Cy7 (BioLegend, cat#157204), anti-CD3-PE (BioLegend, cat#100206), anti-CD8-AF700 (BioLegend, cat#100730), anti- CD69-BV421 (BioLgend, cat#104545, anti-PD-1-PE/Cy7 (BioLegend, cat#109110) and Annexin V-APC (BioLegend, cat#640941), anti-LAG3-PE (BioLegend, cat#125207), anti-TIGIT-Alexa Fluor 700 (R&D system, cat#FAB72671N), anti-TIM3-BV421 (BioLegend, cat#134019).

Techniques: