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