Journal: International Journal of Genomics

Article Title: Integrative Single‐Cell and Bulk Transcriptomics Identifies a Proliferative Malignant State and Supports a Functional Contribution of TROAP in Osteosarcoma

doi: 10.1155/ijog/1407947

Figure Lengend Snippet: Integrated single‐cell landscape of human osteosarcoma. (A) Harmony‐integrated UMAP of 39,063 quality‐controlled cells colored by global cluster. (B) The same embedding colored by patient, showing cross‐patient mixing after integration. (C) Reviewed major‐cell‐type annotations displayed on the integrated UMAP; labels mark lineage centroids. (D) Relative abundance of major cell types in each patient. Colors are consistent between panels C and D.

Article Snippet: HOS and U2OS human osteosarcoma cell lines were obtained from the American Type Culture Collection (ATCC; Manassas, Virginia, United States; HOS, CRL‐1543; U2OS, HTB‐96).

Techniques: Single Cell

Journal: International Journal of Genomics

Article Title: Integrative Single‐Cell and Bulk Transcriptomics Identifies a Proliferative Malignant State and Supports a Functional Contribution of TROAP in Osteosarcoma

doi: 10.1155/ijog/1407947

Figure Lengend Snippet: TROAP depletion reduces clonogenic growth, overall wound closure, migration, and invasion in osteosarcoma cells. (A) Immunoblot analysis of TROAP in HOS and U2OS cells transfected with siNC, siTROAP‐1, or siTROAP‐2; beta‐actin is the loading control. (B) Colony‐formation images and colony counts. (C) Scratch‐wound images at 0 and 24 h and overall wound closure. Because the assay was not proliferation‐matched, it is not interpreted as direct migration evidence. (D) Transwell migration and invasion images and cell counts. Data are mean + /−standard deviation from three independent biological experiments. One‐way ANOVA with Dunnett′s test compared each siTROAP group with siNC; ∗∗∗ p < 0.001.

Article Snippet: HOS and U2OS human osteosarcoma cell lines were obtained from the American Type Culture Collection (ATCC; Manassas, Virginia, United States; HOS, CRL‐1543; U2OS, HTB‐96).

Techniques: Migration, Western Blot, Analysis, Transfection, Control, Colony Assay, Standard Deviation

Journal: The Journal of Biological Chemistry

Article Title: Biosensor-guided discovery of peptide inhibitors targeting the ribosomal protein uS5-PDCD2 chaperone interaction

doi: 10.1016/j.jbc.2026.111415

Figure Lengend Snippet: Inhibition of the uS5-PDCD2 interaction using residues 21 to 50 of uS5. A , Western blot analysis of total extracts (Input, Top ) and anti-Flag purifications (IP:Flag, Bottom ) prepared from HeLa cells that stably expressed Flag-PDCD2 and that were previously transfected with constructs encoding either GFP alone (lane 1), wild-type (WT, lane 2) or F29Y (MT, lane 3) versions of uS5 21-50 -GFP. The blots were analyzed for GFP, Actin, uS5, and Flag-PDCD2. B , quantification of relative uS5 levels copurified from anti-Flag precipitates normalized to Flag-PDCD2 levels. Values were expressed relative to the GFP control vector, which was set to 1.0. The data and error bars represent the average and SD from at least three independent experiments. p -value is indicated, as determined by a one-way ANOVA with Dunnett’s multiple comparisons test. ( C ) U2OS cells that conditionally express Flag-PDCD2 were transfected with either GFP control vector (panels a-d), wild-type (panels e-h) or F29Y (panels i-l) versions of uS5 21-50 -GFP. At the time of transfection, doxycycline was added to the media to induce the expression of Flag-PDCD2. 48 h post-transfection, cells were fixed and simultaneously analysed by direct fluorescence (b, f and j) and immunostaining for Flag-PDCD2 (c, g and k). DNA staining with DAPI shows the nucleus of each cell (a, e and i). Scale bars correspond to 20 μm. D , quantification of nucleus-to-cytoplasmic ratios of Flag-PDCD2 shows a significant decrease in cells that expressed the wild-type version of uS5 21-50 -GFP. More than 60 cells were analyzed for each condition from three independent immunofluorescence experiments. Statistical differences were calculated using a one-way ANOVA with Dunnett’s multiple comparisons test. p -value is indicated. E , Peptide sequences corresponding to residues 21 to 50 of uS5, showing conserved phenylalanines in blue (WT) and substitutions to alanine residues in the mutant in red (MT). F , schematic of the peptide competition experiments using purified uS5-GFP/PDCD2 complex. See text for details. G , Western blot analysis of an uS5-GFP immunoprecipitate that were washed, divided, and treated with increasing concentrations of either wild-type ( Top , WT) or mutant ( Bottom , MT) uS5 21-50 peptides (lanes 2–5) or with no peptide (lane 1). Blots were analyzed for uS5-GFP and endogenous PDCD2. ( H ) Quantification of PDCD2 levels copurified from anti-GFP precipitates normalized to uS5-GFP levels. The values were then set to 1.0 for the control purification in the absence of peptide. Solid lines mark the average binding from three independent replicates, with error bars corresponding to standard deviations. Statistical differences were calculated using a one-way ANOVA with Dunnett’s multiple comparisons test. p -value is indicated.

Article Snippet: Human MOLT-4, HEK293T, HeLa, and U2OS cell lines were acquired from ATCC.

Techniques: Inhibition, Western Blot, Stable Transfection, Transfection, Construct, Control, Plasmid Preparation, Expressing, Fluorescence, Immunostaining, Staining, Immunofluorescence, Mutagenesis, Purification, Binding Assay