Journal: JCI Insight

Article Title: Repression of AXL expression by AP-1/JNK blockage overcomes resistance to PI3Ka therapy

doi: 10.1172/jci.insight.125341

Figure Lengend Snippet: (A) RNA sequencing data obtained from isogenic BYL719 sensitive vs. acquired resistance CAL33 and KYSE180 cells (HNSCC and ESCC, respectively). Presented in the Venn diagram are TF signatures upregulated in BYL719-resistant cells and their overlap with binding sites for TF in the promoter of AXL. (B) WB analysis of CAL33 and KYSE180 BYL719-sensitive vs. -resistant cells showing the expression of TFs (l.e., long exposure; s.e., short exposure). (C) A qPCR analysis comparing the mRNA levels of AXL and c-JUN in CAL33- and KYSE180-sensitive vs.-resistant cells (n > 10). (D) IF images of CAL33- and KYSE180 BYL719–sensitive vs. –resistant cells showing c-JUN (CY-3-labeled) in red, AXL (Alexa488-labeled) in green, and DAPI in blue. Scale bar: 100 μm. (E) Upper panel: WB analysis showing AXL levels after transfection with siRNA for the silencing of c-JUN and c-FOS. Lower panel: qPCR analysis showing AXL mRNA levels in cells transfected with siRNAs for the silencing of c-JUN and c-FOS (n > 6). All WB analysis was assessed in 2–4 independent experiments. All qPCR analysis was assessed in 2–4 independent experiments. One-way ANOVA P value is shown. **P < 0.01.

Article Snippet: For the production of shRNAs cell lines, we created lentiviruses by transfecting HEK293 cells with the viral plasmids psPAX2, pMD2.G, and PLKO with shRNAs — a control scrambled sequence (shCT) or 2 different sequences for the silencing of AXL expression (shAXL1 and shAXL2, MSKCC RNAi core facility identifiers TRCN0000001037 and TRCN0000194771, respectively) using PolyJet transfection reagent (SignaGen, SL100688) according to the manufacturer’s protocol.

Techniques: RNA Sequencing Assay, Binding Assay, Expressing, Labeling, Transfection

Journal: JCI Insight

Article Title: Repression of AXL expression by AP-1/JNK blockage overcomes resistance to PI3Ka therapy

doi: 10.1172/jci.insight.125341

Figure Lengend Snippet: (A) Viability was assessed in cell lines treated with escalating doses of BYL719 for 4 days. Analysis of BYL719 IC50 values in HNSCC and ESCC cells following transfection with siRNAs to silence c-JUN and c-FOS expression (n > 6). (B) Viability was assessed in cell lines treated with BYL719 and SP600125 for 4 days. Analysis of BYL719 IC50 values following JNK inhibition with SP600125 (5 and 10 μM) in HNSCC and ESCC cells (n > 6). (C) WB analysis showing AXL level and AKT/mTOR pathway activation in HNSCC and ESCC cells treated with BYL719 (2 μM), SP600125 (5 and 10 μM), and the combination therapy for 24 hours. (D) qPCR analysis of AXL mRNA levels in cells treated as in with BYL719 (2 μM), SP600125 (10 μM), and combination for 24 hours (n > 6). (E) Viability was assessed in cell lines treated with escalating doses of BYL719 and SP600125 for 4 days. Synergy test for the interaction between BYL719 and SP600125. The synergy test was analyzed using Chalice software (Horizon), and a synergy score was extracted. All WB analysis was assessed in 2–3 independent experiments. All viability experiments were assessed in 2–3 independent experiments. All qPCR experiments were assessed in 2–3 independent experiments. One-way ANOVA P values are shown. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: For the production of shRNAs cell lines, we created lentiviruses by transfecting HEK293 cells with the viral plasmids psPAX2, pMD2.G, and PLKO with shRNAs — a control scrambled sequence (shCT) or 2 different sequences for the silencing of AXL expression (shAXL1 and shAXL2, MSKCC RNAi core facility identifiers TRCN0000001037 and TRCN0000194771, respectively) using PolyJet transfection reagent (SignaGen, SL100688) according to the manufacturer’s protocol.

Techniques: Transfection, Expressing, Inhibition, Activation Assay, Software

Journal: iScience

Article Title: Y RNAs are conserved endogenous RIG-I ligands across RNA virus infection and are targeted by HIV-1

doi: 10.1016/j.isci.2022.104599

Figure Lengend Snippet:

Article Snippet: Polyjet , Signagen , Cat: #SL100688.

Techniques: Strep-tag, Virus, Recombinant, Lysis, Molecular Weight, High Molecular Weight, Modification, Saline, Western Blot, Cell Isolation, Luciferase, Reporter Assay, SYBR Green Assay, Sequencing, In Vitro, CRISPR, Knock-Out, Plasmid Preparation, Control, Software, Real-time Polymerase Chain Reaction

Journal: eLife

Article Title: Single-color, ratiometric biosensors for detecting signaling activities in live cells

doi: 10.7554/eLife.35458

Figure Lengend Snippet:

Article Snippet: Commerical Kit , Polyjet , SignaGen , SL100688.

Techniques: Recombinant, Software