Viewrna red stain kit

About 4 μm-thick formalin-fixed paraffin-embedded sections of IDHWGB tissues (n = 3) selected from the original cohort of six patients were used for mRNA CISH staining, using the Leica Bond Rx autostainer and detected using the ViewRNA red stain kit (Affymetrix, Santa Clara, CA, USA). Probes for cathepsin B (NM_001908), cathepsin D (NM_001909), and cathepsin G (NM_001911) were obtained from Affymetrix. Human placenta, human breast tissue, and mouse bone marrow were used as positive controls for cathepsin B, cathepsin D, and cathepsin G, respectively. Negative controls were exhibited on sections of GBM tissue using a probe for Bacillus (cat# VF1-11712, Affymetrix).

Representative 4 μm-thick, formalin-fixed, paraffin-embedded sections of three MDBMSCC samples from the original cohort of six patients used for DAB IHC staining were used for mRNA colorimetric in situ hybridization (CISH). Staining was done on the Leica Bond Rx auto-stainer and detected using the ViewRNA red stain kit (Affymetrix, Santa Clara, CA, USA), as previously described (35 (link)). The probes used for NANOG (NM_024865), SOX2 (NM_003106), SALL4 (NM_020436), STAT3 (NM_003150), and OCT4 (NM_002701) were obtained from Affymetrix. Positive controls used were human IH for SALL4 and human seminoma for NANOG, SOX2, OCT4, and STAT3. A negative control for the primary antibody was done on a sample of MDBMSCC from the CISH cohort by omitting the probe.

4 μm thick formalin-fixed paraffin-embedded sections of six samples of MDOTSCC from the original cohort of nine patients used for DAB IHC staining were used for CISH. Staining was carried out on the Leica Bond Rx auto-stainer and detected using the ViewRNA red stain kit (Affymetrix, Santa Clara, CA, USA), as previously described (16 (link)). The probes used for cathepsin B (cat# VA1-12282), cathepsin D (cat# VA1-12281), cathepsin G (cat# VA1-21016), and Bacillus (cat# VF1-11712, Affymetrix) as a negative control. Positive controls used were the same for DAB IHC staining.