diff-quick stain  (PanReac AppliChem)

Journal: Cancer Cell International

Article Title: Inhibition of RBM23 induces ferroptosis in colon cancer cells via c-Myc regulation

doi: 10.1186/s12935-026-04262-x

Figure Lengend Snippet: RBM23 knockdown inhibits colon cancer cell growth and survival (A) Protein was extracted from the normal colon cell lines CCD-18Co and colon cancer cell lines (HCT116, HT29, LOVO, LS513, DLD-1, COLO205, HCT15) and quantified in equal amounts, and then the protein expression of RBM23 was detected by western blotting. (B) The mRNA expression level of RBM23 was measured by qRT-PCR in the normal colon cell line CCD-18Co and colon cancer cell lines (DLD-1, HCT15, HT29, HCT116). GAPDH was used as an internal control for normalization. (C) Western blot analysis was performed to measure the expression levels of c-Myc in HCT15 cells treated with control siRNA or RBM23 siRNA for 72 h. (D) HCT116 cells were treated with control siRNA or RBM23 siRNA #1, #2, #3 for 72 h. Protein expression levels of various factors were then analyzed by western blotting. (E) The effect of RBM23 deficiency on HCT116, a colon cancer cell line. HCT116 was transfected with 80 nM RBM23 siRNA and control siRNA, and cell viability was examined at 0 and 72 h of treatment. Cell viability was measured by CCK8 assay and optical density was measured by microplate reader at 450 nm. Data were normalized to the control siRNA group at 0 h (set as 100%). (F) The effect of RBM23 knockdown in colon cancer cells, HCT116, on cell proliferation. HCT116 cells were transfected with RBM23 siRNA (100 nM) and incubated for one week, and colonies were stained with Diff Quick solution. The colonies were counted and quantified. (G) Western blotting was performed to detect proteins extracted from HCT116 cells fractionated into nucleus and cytoplasm. Proteins from HCT116 cells transfected with RBM23 siRNA (100 nM) for 72 h were extracted and separated by NE-PER nuclear and cytoplasmic extraction kit into proteins extracted from the nucleus and cytoplasm. The proteins were then detected using western blotting. Student’s t -test: ***P < 0.001

Article Snippet: The plates were then incubated at 37 °C, 5% CO 2 for 1 week and the colonies were fixed and stained using Diff-Quick Kit (Sysmex Corporation, Kobe, Hyogo, Japan) and the number of colonies was counted.

Techniques: Knockdown, Expressing, Western Blot, Quantitative RT-PCR, Control, Transfection, CCK-8 Assay, Incubation, Staining, Diff-Quik, Extraction