Journal: Frontiers in Oncology

Article Title: Identification and comprehensive analysis of an immune-related gene prognostic model for indicating tumor immune microenvironment features in soft tissue sarcoma

doi: 10.3389/fonc.2025.1609501

Figure Lengend Snippet: CALR is overexpressed in osteosarcoma and modulates cellular phenotypes. (A) qRT-PCR analysis of CALR mRNA levels in osteosarcoma cell lines (MG-63, 143B, HOS) compared to human normal osteoblasts (hFOB1.19). (B) Western blot showing endogenous CALR protein expression in untreated osteosarcoma cells (Saos-2, MG-63, 143B, U2OS) and CALR-overexpressing 143B cells (OE-CALR). (C) Immunofluorescence staining of CALR (TRITC, red) and nuclei (DAPI, blue) in 143B cells under overexpression (OE-CALR), knockdown (sh-CALR), and negative control (NC) conditions. Fluorescence intensity quantification is shown below. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Cells on coverslips were fixed with 4% PFA, permeabilized with 0.2% Triton X-100, blocked with 3% BSA, and incubated with anti-CALR antibody (#10208-1-AP, 1:200, Proteintech) overnight at 4°C, followed by TRITC-conjugated secondary antibody (1:500) and DAPI (1 μg/mL) at RT.

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Immunofluorescence, Staining, Over Expression, Knockdown, Negative Control, Fluorescence