crl 2539  (Procell Inc)

Journal: PLOS ONE

Article Title: A novel TREX1 inhibitor, VB-85680, upregulates cellular interferon responses

doi: 10.1371/journal.pone.0305962

Figure Lengend Snippet: A) TREX1 Western blot demonstrating overexpression of both wild-type mouse TREX1 and mouse TREX1 carrying the D18N inactivating mutation. Endogenous mouse TREX1 present in a J774 cellular lysate is shown as reference to confirm the size of the upper band as full-length mouse TREX1. α-tubulin was used as a loading control. B–D) Exonuclease assays performed with HEK293T mouse TREX1 overexpression or empty vector lysates demonstrate the effects of VB-85680 and VB-85662 on B) wild-type mouse TREX1 (5 ng/μL total protein), C) Catalytically inactive mouse TREX1 D18N (100 ng/μL total protein) and, D) endogenous exonuclease activity (100 ng/μL total protein). E) Western blot for endogenous levels of cGAS,STING and TREX1 in both 4T1 and THP1-dual cells. F) Endogenous exonuclease activity in 4T1 cell lysates is inhibited by VB-85680, but not by VB-85662. Compounds were titrated in the presence of 6 μg 4T1 cytosolic lysate using the 3’ to 5’ Exonuclease Activity Assay from BioVision. G) Inhibition of endogenous human TREX1 exonuclease activity by VB-85680 in THP1-Dual™ and THP1-Dual™ TREX1 knockout cell lysates. The exonuclease assay was carried out as described in 1E . Error bars for all experiments represent +/- SD.

Article Snippet: Briefly, THP1-Dual™ (Invivogen thpd-nfis), THP1-Dual™ TREX1 KO (Invivogen thpd-nfis) or 4T1 (ATCC CRL-2539) cells were suspended at a density of 1 million cells/mL in the lysis buffer provided.

Techniques: Western Blot, Over Expression, Mutagenesis, Control, Plasmid Preparation, Activity Assay, Inhibition, Knock-Out

Journal: Nature Communications

Article Title: Blocker-SELEX: a structure-guided strategy for developing inhibitory aptamers disrupting undruggable transcription factor interactions

doi: 10.1038/s41467-024-51197-w

Figure Lengend Snippet: a CoIP assays of WMiApt-1 in blocking the interaction between WDR5 and MYC (biological replicates n = 2). Source data are provided as a Source Data file. b Protein levels of MYC and WDR5 of three human-derived cell lines. c Anti-proliferation effects of WMiApt-1 in three human cell lines The result is determined from three biological replicates, each containing three technical replicates (biological replicates n = 3, mean ± SD, Two-sided Student’s t -test). The 95% confidence intervalare −56.79 to −36.85, −23.20 to −2.945 and −8.267 to −0.7118. Source data are provided as a Source Data file. Source data are provided as a Source Data file. d Schematic diagram illustrating the anti-tumorigenesis experiments in-vivo. Icons from Canva. e Tumor volumes of 4T1 cells treated with WMiApt-1 or control on the fifth day post-implantation (biological replicates n = 5, mean ± SD, Two-sided Student’s t -test). The 95% confidence interval is −8.267 to −0.7118. Source data are provided as a Source Data file.

Article Snippet: MDA-MB-468 (HTB-132), HEK293T (ACS-4500), SKNBE2 (CRL-2271), 4T1 (CRL-2539) cell lines were purchased from ATCC.

Techniques: Blocking Assay, Derivative Assay, In Vivo, Control

Journal: Nature Communications

Article Title: Reprogramming the tumor immune microenvironment using engineered dual-drug loaded Salmonella

doi: 10.1038/s41467-024-50950-5

Figure Lengend Snippet: a Experimental scheme. Briefly, 4T1 cells were implanted into the mammary fat pad of BALB/c mice. When tumors reached approximately 140 mm 3 , mice received bacteria through i.v injection (day 0). Doxy (+) and CSF-1R antibody (αCSF-1R) were administered as shown. Pimary tumors and metastatic lungs were collected on day 15 and 35, respectively ( b , c ) and flow cytometry was performed on day 9 ( d , e ). b Average growth curves of 4T1 primary tumors ( n = 6 mice examined from two independent experimental replicates; * P = 0.0144, **** P < 0.0001; two-way ANOVA with Tukey’s multiple comparisons test). c Individual images (left) and metastatic nodule count (right) of lungs ( n = 6 exami n ed from two independent experimental replicates; **** P < 0.0001; ns not significant; unpaired two-tailed t -test). d Flow cytome t ry gating of M1 (F4/80 + CD86 + ) and M2 (F4/80 + CD206 + ) macrophages . e Ratio of M1/M2 macrophages in lungs (left) and tumors (right) ( n = 6 mice examined from two independent experimental replicates; **** P < 0.0001; ns, not significant; unpaired two-tailed t -test). f In vivo and ex vivo bioluminescent imaging of MDA-MB-231-FLuc-GFP lung metastasis. Left, images of the mice on day 0 (pre-treatment) and on day 14 (post-treatment) and the excised lungs on day 14; right, quantitation of bioluminescence levels in the lungs. g Frequency of macrophages phagocytosing tumor cells (F4/80 + GFP + ) in the metastatic lungs of ( f ) ( n = 3 mice/group, from one experiment, unpaired two-tailed t -test). h Experimental scheme. i Average growth curves of subcutaneous tumors ( n = 7 mice examined from two independent experimental replicates; **** P < 0.0001; two-way ANOVA with Tukey’s multiple comparisons test). j Kaplan-Meier survival curves ( n = 7 mice examined from two independent experimental replicates; **** P < 0.0001; Log-rank (Mantel-Cox) test). k Metastatic scores of livers. Metastatic nodules on the liver surface were counted at the experimental end point. The scores were classified as follows: 0, 0%; 1, <20%; 2, 20%–50%; 3, >50% ( n = 7 mice examined from two independent experimental replicates; **** P < 0.0001; unpaired two-tailed t -test).

Article Snippet: CT26 (mouse colon carcinoma, CRL-2638), 4T1 (mouse breast cancer, CRL-2539), and HEK293T (human epithelial cell, CRL-3216) cell lines were purchased from the American Type Culture Collection (ATCC, USA).

Techniques: Bacteria, Injection, Flow Cytometry, Two Tailed Test, In Vivo, Ex Vivo, Imaging, Quantitation Assay