Journal: Frontiers in Immunology

Article Title: A tumor immune microenvironment gene expression signature for predicting prognosis, immunotherapy efficacy, and drug candidates in triple-negative breast cancer

doi: 10.3389/fimmu.2025.1676768

Figure Lengend Snippet: The immune microenvironment–reprogramming function of NCD is mediated by JAK2 inhibition. (A) Volcano plot of DEGs in MDA-MB-231 cells treated with or without 5 μM NCD. (B) KEGG enrichment analysis of DEGs. (C) Western blot of JAK2, p-JAK2, STAT3, and p-STAT3 in MDA-MB-231 cells treated with increasing doses of NCD (n=3). (D) Quantification of JAK2, p-JAK2, STAT3, and p-STAT3 proteins corresponding to (C) (n = 3). (E) Western blot of JAK2, p-JAK2, STAT3, and p-STAT3 in 4T1 cells treated with increasing doses of NCD (n=3). (F) Quantification of JAK2, p-JAK2, STAT3, and p-STAT3 proteins corresponding to (E) (n = 3). (G) Protein expression levels of JAK2 after knockdown of JAK2 in MDA-MB-231 cell line (n=3). (H) Quantification of JAK2 protein levels corresponding to (G) (n=3). (I) mRNA expression levels of JAK2 , CXCL10 , CXCL11 , EBI3 , FLT3LG after knockdown of JAK2 in MDA-MB-231 cell (n=3). (J) Protein stability of JAK2 under thermal gradient with or without NCD (n=3). (K) CETSA Melting Curve (n=3). (L) SPR sensorgrams of JAK2 binding to NCD. (M) Affinity analysis and KD calculation of JAK2-NCD interaction. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Membranes were blocked and then incubated with the following primary antibodies: Stat3 (124H6) Mouse mAb, Phospho-Stat3 (Tyr705) (D3A7) XP ® Rabbit mAb, Jak2 (D2E12) XP ® Rabbit mAb, Phospho-Jak2 (Tyr1007/1008) Antibody (all from Cell Signaling Technology), and Alpha Tubulin Monoclonal antibody (Proteintech).

Techniques: Inhibition, Western Blot, Expressing, Knockdown, Binding Assay

Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: Ursolic acid induces apoptosis and disrupts host-parasite interactions in Theileria annulata -infected cells

doi: 10.1016/j.ijpddr.2025.100593

Figure Lengend Snippet: UA Exerts Multifaceted Effects on Host and Parasite in TA Cells. A. Representative chromatogram of control and B. UA treated TA cells from mass spectrometry analysis. C. and D. Represents Pearson correlation coefficient analysis between two replicates of control and UA treated sample. E. Representative heatmap of Pearson correlation coefficient analysis between all replicates of control and UA treated sample. F. Bar graph represents the significantly affected biological pathways in host. G. Bar graph represents the significantly affected biological pathways in parasite. Analysis was done using Perseus and FunRich (Functional Enrichment Analysis Tool). H. Heatmap represents the ANOVA significant genes affected in host and I. parasite between all four control and UA treated TA cells. J. Western blotting shows the expression of phosphorylated S/T proteins and H3K9ME3 from whole cell lysate of TA cells after UA treatment for 6, 12 and 24 h. K. Western blotting shows the expression of JAK2 and c-Jun from whole cell lysate of TA cells after UA treatment for 6 and 12 h. Numerical values represent the normalized values from loading control β-actin.

Article Snippet: Primary antibodies against cleaved caspase-3 (CST 9661), cleaved caspase-8 (CST 8592), cleaved caspase-9 (CST, 20750), total caspase-3 (CST 9662), T. annulata surface protein (TaSP) peptide, c-Jun (CST 9165), JAK2 (CST 3230), and phospho-serine/threonine (CST 9631) were used at 1:1000 dilution.

Techniques: Control, Mass Spectrometry, Functional Assay, Western Blot, Expressing

Journal: Scientific Reports

Article Title: A tryptophol-containing emulgel ameliorates imiquimod-induced mice psoriasis

doi: 10.1038/s41598-025-04431-4

Figure Lengend Snippet: The TOH-containing emulgel alleviated activation of the JAK2/STAT3 pathway in inflammation and cell proliferation. ( a ) Immunohistochemical staining for JAK2, pJAK2, STAT3, and pSTAT3 on the skin section of all groups was performed. Representative images of skin sections were taken at ×40 magnification (scale bars: 50 μm). ( b , c ) Quantification of JAK2, and STAT3 positive cells in all groups (n = 4). ( d , e ) The mRNA expression of JAK2 and STAT3 . ( f ) Protein expression of JAK2, pJAK2, STAT3 and pSTAT3 evaluated by western blot. ( g , h ) Relative protein expression levels of pJAK2 and pSTAT3 normalized to total JAK2 and STAT3, respectively (n = 4). ( j , k ) Quantification of pJAK2, and pSTAT3 positive cells in all groups (n = 4). ( l , m ) The mRNA expression of BCL2 and CCND1 were analyzed by RT-qPCR (n = 4). The data are presented as the means ± SD. Statistical significance was determined by one-way ANOVA. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: A rabbit monoclonal anti-β-actin (13E5) antibody (#4970S), rabbit monoclonal anti-phosphor-Stat3 (Tyr705) antibody (D3A7), rabbit monoclonal anti-Jak2 antibody (D2E12) were purchased from Cell Signaling Technology (Danvers, Massachusetts, USA).

Techniques: Activation Assay, Immunohistochemical staining, Staining, Expressing, Western Blot, Quantitative RT-PCR