rabbit anti p70 s6k thr389  (Cell Signaling Technology Inc)

Journal: Signal Transduction and Targeted Therapy

Article Title: Breast cancer-associated fibroblast-derived exosomal miR-1290 promotes triple-negative breast cancer cell proliferation

doi: 10.1038/s41392-026-02922-y

Figure Lengend Snippet: miR-1290 activates mTORC1 signaling through the downregulation of TBC1D4. a Gene Ontology analysis of commonly downregulated genes in miR-1290-overexpressing and TBC1D4-knockdown BT-549 cells (log2(FPKM) + 1 ratio < 0.95 versus the respective controls). b Representative images of phospho-kinase arrays performed using lysates from BT-549 cells transfected with the miR-1290 mimic or mimic-NC and siTBC1D4 or siCont (left). The relative intensity of p-P70S6K is shown in the graph (right). c Expression of TSC1 in BT-549 cells treated as indicated. d Expression levels of TSC1 in normal ( n = 114) and breast cancer ( n = 1097) tissues from the TCGA database (*** p < 0.001, Mann‒Whitney U test). e Analysis of the correlation between TBC1D4 and TSC1 expression in breast cancer tissues from the TCGA database ( r = 0.251, p < 0.001). f Representative immunocytochemistry images showing the localization and expression of TBC1D4, TSC1, and p-P70S6K in BT-549 and Hs578T cells transfected with the miR-1290 mimic or mimic-NC and with siTBC1D4 or siCont. The nuclei were counterstained with DAPI. Scale bar, 150 μm. g Western blot analysis of the indicated proteins in BT-549 and Hs578T cells transfected with the miR-1290 mimic or mimic-NC and with siTBC1D4 or siCont. ACTB was used as a loading control

Article Snippet: The primary antibodies used were as follows: TBC1D4 (1:100; Cell Signaling Technology, 2670S), TSC1 (1:200; Proteintech, 29906-1-AP), phospho-P70S6K (1:200; ABclonal, AP0564), and Ki67 (1:200; Abcam, ab16667).

Techniques: Analysis, Knockdown, Transfection, Expressing, Immunocytochemistry, Western Blot, Control

Journal: Signal Transduction and Targeted Therapy

Article Title: Breast cancer-associated fibroblast-derived exosomal miR-1290 promotes triple-negative breast cancer cell proliferation

doi: 10.1038/s41392-026-02922-y

Figure Lengend Snippet: Verification of the loss of function of miR-1290 in TNBC cell lines. a Crystal violet staining of BT-549 and Hs578T cells transfected with anta-NC or anta-1290 (left) and quantification of cell numbers (right). Scale bar, 500 μm. b Growth analysis by a CCK-8 assay in BT-549 and Hs578T cells transfected with anta-NC or anta-1290. c qRT‒PCR analysis of miR-1290, TBC1D4, and TSC1 expression in BT-549 and Hs578T cells transfected with anta-NC or anta-1290. d Western blot analysis of TBC1D4, P70S6K, p-P70S6K, Ki67, and ACTB in BT-549 and Hs578T cells transfected with anta-NC or anta-1290. ACTB was used as a loading control. e Representative immunocytochemistry images showing the expression of TBC1D4, TSC1, p-P70S6K, and Ki67 in BT-549 and Hs578T cells transfected with anta-NC or anta-1290. The nuclei were counterstained with DAPI. Scale bar, 150 μm. The quantification of the fluorescence intensity per cell is shown below each panel. The data are shown as the means ± SDs of three independent experiments, and statistical significance was determined using Student’s t tests (* p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: The primary antibodies used were as follows: TBC1D4 (1:100; Cell Signaling Technology, 2670S), TSC1 (1:200; Proteintech, 29906-1-AP), phospho-P70S6K (1:200; ABclonal, AP0564), and Ki67 (1:200; Abcam, ab16667).

Techniques: Staining, Transfection, Analysis, CCK-8 Assay, Expressing, Western Blot, Control, Immunocytochemistry, Fluorescence

Journal: Signal Transduction and Targeted Therapy

Article Title: Breast cancer-associated fibroblast-derived exosomal miR-1290 promotes triple-negative breast cancer cell proliferation

doi: 10.1038/s41392-026-02922-y

Figure Lengend Snippet: Everolimus suppresses miR-1290-mediated tumor growth in vivo. a Growth curves of xenograft tumors from the indicated treatment groups ( n = 4 per group). b Representative images of xenograft tumors from the indicated treatment groups. c Tumor weight measurements at the endpoint of the xenograft experiment ( n = 4 per group). d Representative H&E staining images of tumor sections. Scale bar, 200 μm. e Immunohistochemical analysis of TBC1D4, TSC1, p-P70S6K, and Ki67 expression in tumor sections from the indicated treatment groups (left) and quantification of staining intensity (right) ( n = 4 per group). Scale bar, 200 μm. The data are shown as the means ± SDs, and statistical significance was determined using two-way repeated-measures ANOVA with Tukey’s multiple comparisons test for the tumor growth curves ( a ) and two-way ANOVA with Tukey’s multiple comparisons test for the remaining analyses ( c , e ) (* p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: The primary antibodies used were as follows: TBC1D4 (1:100; Cell Signaling Technology, 2670S), TSC1 (1:200; Proteintech, 29906-1-AP), phospho-P70S6K (1:200; ABclonal, AP0564), and Ki67 (1:200; Abcam, ab16667).

Techniques: In Vivo, Xenograft Assay, Staining, Immunohistochemical staining, Analysis, Expressing

Journal: Tumour Virus Research

Article Title: Oncolytic virus hijacks GOT1 and pyrimidinosomes to fuel pyrimidine synthesis for replication in tumor cells

doi: 10.1016/j.tvr.2026.200342

Figure Lengend Snippet: | The MAS-transported NAD + /NADH regulates CAD phosphorylation activity and NDV replication through S6K. (A) Schematic diagram showing the potential involvement of mTOR-S6K in regulating CAD phosphorylation through MAS-mediated NAD + /NADH translocation. (B-D) A549 cells were either mock-infected or infected with NDV (MOI = 1) at 6, 12, 18 and 24 hpi. Protein levels of p-S6K (T389), S6K, p-S6 (S235/236), S6, p-CAD (S1859), CAD, NP and β-actin were analyzed by WB at 12 hpi (B). Grayscale quantification of p-S6K/S6K, p-S6/S6 and p-CAD/CAD ratios from WB data (C). Intracellular NAD + /NADH ratio were measured (D). (E-H) A549 cells were infected with NDV (MOI = 1), treated with AOA, supplemented with Pyr or Asp. Protein levels of p-S6K, S6K, p-S6, S6, p-CAD, CAD, NP, and β-actin were analyzed by WB at 12 hpi (E). Grayscale quantification of p-S6K/S6K, p-S6/S6 and p-CAD/CAD ratios from WB data (F). Intracellular NAD + /NADH ratio were measured (G). Extracellular virus yields were detected (H). (I-L) A549 cells were infected with NDV (MOI = 1), treated with AOA, supplemented with Pyr, and exposed to Torin 1. Protein levels of p-S6K, S6K, p-S6, S6, p-CAD, CAD, NP, and β-actin were analyzed by WB at 12 hpi (I). Grayscale quantification of p-S6K/S6K, p-S6/S6 and p-CAD/CAD ratios from WB data (J). Intracellular NAD + /NADH ratio were measured (K). Extracellular virus yields were detected (L). (M − P) A549 cells were infected with NDV (MOI = 1), treated with AOA, and supplemented with Duro. Protein levels of p-S6K, S6K, p-S6, S6, p-CAD, CAD, NP, and β-actin were analyzed by WB at 12 hpi (M). Grayscale quantification of p-S6K/S6K, p-S6/S6 and p-CAD/CAD ratios from WB data (N). Intracellular NAD + /NADH ratio were measured (O). Extracellular virus yields were detected (P). Data are presented as means from three independent experiments, ns, p > 0.05, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: The antibodies used for immunoblotting in this study included monoclonal or polyclonal: anti-β-actin (Proteintech, Cat# 66009-1-Ig), anti-NP (Prepared in our laboratory), anti-S6 ribosomal protein (Cell Signaling Technology [CST], Cat# 2217), anti-phospho-S6 ribosomal protein (S235/236) (CST, Cat# 4858), anti-phospho-p70 S6 Kinase 1 (T389) (ABclonal, Cat# AP0564), anti-p70 S6 Kinase 1 (ABclonal, Cat# A2190), anti-CAD (CST, Cat# 11933), anti-phospho-CAD (S1859) (CST, Cat# 70307), anti-GOT1 (Proteintech, Cat# 14886-1-AP), anti-GOT2 (Proteintech, Cat# 14800-1-AP). anti-UMPS (Proteintech, Cat# 14830-1-AP), anti-DHODH (Proteintech, Cat# 14877-1-AP) and anti-Tom20 (Abcam, Cat# ab283317).

Techniques: Phospho-proteomics, Activity Assay, Translocation Assay, Infection, Virus