Journal: bioRxiv

Article Title: Salinomycin disturbs Golgi apparatus function and specifically affects cells in epithelial-to-mesenchymal transition

doi: 10.1101/2022.08.31.506024

Figure Lengend Snippet: (A) Western blot analysis of membrane proteins N- and E-cadherin; epidermal growth factor receptor and transferrin receptor (EGFR and TFRC) in HMLE-shEcad and HMLE-shGFP breast cells (A) or three different breast tumor cell lines (B) that were treated for 24 hours with indicated concentrations of DMSO (Mock), salinomycin (Sal), nigericin (Nig), monensin (Mon) or paclitaxel (PTX). Naphtol blue staining of transferred proteins was used to evaluate equal loading; bloting of E-cadherin in (B) was performed after N-cadherin on the same membrane and it is showing in SUM159 cells, protein bands denoted by #. (C) Western blot analysis of proN-cadherin with specific N-cadherin prodomain antibody in HMLE-Twist cells after treatment with Sal for 24h. (D) Western blot analysis of proN- and N-cadherin in HMLE-Twist cells after treatment with PNGase F. (E) HMLE-Twist and HMLE-pBp cells were treated with DMSO (mock), salinomycin (Sal) or monensin (Mon) for 24 hours and media above cells was subjected to ELISA analysis of secreted IL-8. Data was normalized to mock-treated sample.

Article Snippet: Primary antibodies used: E-cadherin (1:500, Becton-Dickinson, 610181), N-cadherin (1:500, Becton-Dickinson, 610920), proN-cadherin (1:500, R&D Systems, AF1388), EGFR (1:1000, Cell Signaling Technology, 4267), TFRC (1:500, Thermo Fisher Scientific, 13-6800), eIF2α (1:1000, Cell Signaling Technology, 9722), p-eIF2α (1:1000, Cell Signaling Technologies, 3597), CREB-2 (1:100, SCBT, sc-200).

Techniques: Western Blot, Membrane, Staining, Enzyme-linked Immunosorbent Assay

Journal: Cell reports

Article Title: Germline-specific RNA helicase DDX4 forms cytoplasmic granules in cancer cells and promotes tumor growth.

doi: 10.1016/j.celrep.2024.114430

Figure Lengend Snippet: Figure 5. Transcriptome is misregulated in DDX4-null PC3 cells and DDX4-null PC3 xenograft tumors (A) Hierarchical heatmaps show differentially expressed (|log2FC| R 1, adjusted p value %0.05) genes in DDX4-null vs. WT PC3 cells (left) and xenograft tumors (right). (B) Venn diagram shows the overlap between the genes upregulated in DDX4-null vs. WT PC3 xenograft tumors and cells. (C) Volcano plot shows the differential expression of tumor suppressor genes in DDX4-null vs. WT PC3 tumors, with significantly upregulated (green) and downregulated (orange) genes labeled. (D) Bar chart shows the log2FC (DDX4-null vs. WT tumors) of individually selected genes. (E) Bar chart shows the validation of the selected differentially expressed genes in DDX4-null vs. WT PC3 tumors (number of biological replicates is 3) by qRT-PCR (ADAM12: p = 0.0112, CDH6: p = 0.0161, CDH7: p = 0.0298, CEACAM1: p = 0.0468, SFRP1: p = 0.0293, and CLEC2B: p = 0.0105, Mann-Whitney U test, 2-tailed). (F) Western blotting image of CDH6, CDH1, and CDH2 protein expression in DDX4-null and WT PC3 tumors (3 biological replicates each). b-Actin was used as the loading control. Bar chart shows the quantification of the CDH1, CDH2, and CDH6 protein levels normalized by b-actin signal. CDH1: p = 0.0055, CDH6: p = 0.0004 (Mann-Whitney U test, 2-tailed, 3 biological replicates each). Data are represented as mean ± SEM. See also Figures S3 and S4 and Table S1.

Article Snippet: Primary antibodies used for immunoprecipitation andwestern blotting: DDX4 (ProteinTech, 510421-1-AP), CDH1 (Cell signaling technology, 14472S), CDH2 (R&D systems, MAB13881) and CDH6 (Cell signaling technology, 48111S) were used for western blotting.

Techniques: Quantitative Proteomics, Labeling, Biomarker Discovery, Quantitative RT-PCR, MANN-WHITNEY, Western Blot, Expressing, Control