Journal: Nature Communications

Article Title: Co-targeting JAK1/STAT6/GAS6/TAM signaling improves chemotherapy efficacy in Ewing sarcoma

doi: 10.1038/s41467-024-49667-2

Figure Lengend Snippet: a A cartoon illustration of the experimental procedure for the phospho-profiling experiment generated by Biorender. Please refer to the Method section for details. b The signals from the phospho-profiling assay. Increased phosphorylation signals are boxed and labeled with letters. c , d Quantifications of the phospho-profiling signals in ( b ) and representative signals for each letter from ( b ) and grouped in ( d ). Error bars were calculated as mean ± SD, n = 2 (technical duplicates). e , f Immunoblot (IB) analysis of whole cell lysates (WCL) from A673 cells ( e ) or MHH-ES-1 cells ( f ) treated with indicated doses of SN-38 for 24 h. g , h IB analysis of WCL from A673 cells ( g ) or MHH-ES-1 cells ( h ) treated with indicated doses of TMZ (temozolomide) for 24 h. i , j IB analysis of WCL from A673 cells ( i ) or MHH-ES-1 cells ( j ) treated with indicated doses of etoposide for 24 h. k , l IB analysis of WCL from A673 cells ( k ) or MHH-ES-1 cells ( l ) treated with indicated doses of doxorubicin for 24 h. m Tumor volume measurements at indicated days post-injection with temozolomide injected at the indicated days. Temozolomide (50 mg/kg) were injected via IP. Error bars were calculated as mean ± SD, n = 6 tumors. p values are calculated as indicated (two-way ANOVA followed by Tukey multiple comparison test). n Isolated tumors from ( m ) and weighed in ( o ). Error bars were calculated as mean ± SD, n = 6 tumors. p values are calculated as indicated (two-tailed student’s t -test). p IB analysis of WCL derived from dissected tumors with or without Temozolomide treatment. The number indicates each tumor obtained from animals.

Article Snippet: MHH-ES-1, A673, and SK-N-MC cells have been recently authenticated by the Davis Lab by STR analyses (Genetica).

Techniques: Generated, Phospho-proteomics, Labeling, Western Blot, Injection, Comparison, Isolation, Two Tailed Test, Derivative Assay

Journal: Nature Communications

Article Title: Co-targeting JAK1/STAT6/GAS6/TAM signaling improves chemotherapy efficacy in Ewing sarcoma

doi: 10.1038/s41467-024-49667-2

Figure Lengend Snippet: a , b Representative heatmaps for cell viability in MHH-ES-1 cells treated with indicated doses of two compounds (as indicated in X and Y axes, respectively) for 2 days. The color scale bar indicates % of survived cells, where 1.0 represents 100%. c IB analysis of WCL derived from A673 cells treated with indicated compounds for 2 h followed by the addition of 1 μM SN-38 overnight before cell collection. d – f IB analysis of WCL derived from MHH-ES-1 cells depleted of MERTK ( d ), AXL ( e ), or TYRO3 ( f ). Where indicated, MHH-ES-1 cells were infected with indicated sgRNA viruses and selected in 1 μg/mL puromycin to eliminate non-infected cells for 72 h before cell collection . g – l Cell viability assays using indicated MHH-ES-1 cells treated with indicated doses of chemotherapeutic agents for 48 h. Error bars were calculated as mean ± SD, n = 3 (experimental triplicates). * p < 0.05 represents differences between the experimental groups compared to the control group (one-way ANOVA test). * p value of each point ( g : sgMERTK/sgctrl: 0.0012&0.0004 (6 nM); 0.0012 & <0.0001 (10 nM); 0.0038&0.0007 (30 nM); h : sgAXL/sgctrl: 0.002&0.0002 (6 nM), 0.0007&0.0013 (10 nM), 0.0004&0.0005 (30 nM); i : sgTYRO3/sgctrl: 0.0004&0.0003 (6 nM), <0.0001& < 0.0001 (10 nM), <0.0001& < 0.0001 (30 nM); j: sgAXL/sgctrl: <0.0001&0.0001 (10 nM), 0.001&0.0006 (30 nM), 0.0127&0.0147 (100 nM), 0.001&0.0008 (300 nM); k: sgTYRO3/sgctrl: 0.0004&0.0003 (10 nM), 0.0066&0.0236 (30 nM); l: sgMERTK/sgctrl: 0.0026&0.0005 (10 nM), 0.0001 & <0.0001 (30 nM), 0.0009 & 0.0006 (100 nM). WB data presented in this figure are representative data from experimental duplicates.

Article Snippet: MHH-ES-1, A673, and SK-N-MC cells have been recently authenticated by the Davis Lab by STR analyses (Genetica).

Techniques: Derivative Assay, Infection, Control