anti id1  (Cell Signaling Technology Inc)

Journal: Frontiers in Immunology

Article Title: Generation and characterization of human induced pluripotent stem cells from neuropathologically confirmed multiple system atrophy patient-derived fibroblasts

doi: 10.3389/fimmu.2026.1641981

Figure Lengend Snippet: Assessment of the cell line identity and the genomic integrity of the six MSA patient-derived iPSCs. (A) Short tandem repeat profiling provided the number of repeating units of 16 polymorphic microsatellite loci, to compare the donor fibroblast cells (reference) and the generated iPSCs and verify their origin. (B) G-banding revealed that the six iPSC lines showed a normal diploid karyotype (n = 46) and matched the sex of the donor patient. (C) Digital PCR allowed us to screen whether specific genomic regions accumulated duplications that may give a strong selective survival or growth advantage to iPSCs. Example of the scatter digital PCR plots displaying simultaneously (1) FAM signals, shown in purple and green to detect the target gene ( ID1 , NCAPD2 , PITX1 , RPS6KB1 , SOAT1 , or STS ), which are located in the 6 most recurrent abnormal regions; and (2) VIM signals, shown in orange and green to detect the reference gene, RPP30 . The ratio between the number of copies of the target gene and of the reference gene was used to determine the normal 2-copy number or copy number variations. The plots were obtained through QuantStudio 3D Digital PCR system. In all cases, we obtained 2-copy number (D) Quantitative Real Time RT-PCR was used for confirming the absence of the Sendai virus in the iPSC lines after 8 passages, through four primer sets (SeV, KOS, Klf4 and c-Myc). Box and Whisker plots depict in log10 scale the gene expression relative to GAPDH gene and normalized to the 7 days post-transduction fibroblasts, calculated by the 2(-ΔΔC(T)) method. In all iPSC clones, the expression levels of the SeV RNA were very low, reaching the limit of detection. Data are shown as the mean ± s.d. n = 3.

Article Snippet: ID1 , Hs01892845_cn , FAM , 20q11 , hg38|31605682-31605761 , 79 bp , MseI, HindIII, CviQI.

Techniques: Derivative Assay, Generated, Digital PCR, Quantitative RT-PCR, Virus, Whisker Assay, Gene Expression, Transduction, Clone Assay, Expressing

Journal: bioRxiv

Article Title: Increases in BCL2L1 and ID1 dosage synergistically drive fate bias and competitive advantage in human pluripotent stem cells

doi: 10.64898/2026.03.26.714405

Figure Lengend Snippet: Effects of BCL2L1 and ID1 overexpression on hESC differentiation. (B) Quantitative RT-PCR analysis of BCL2L1 and ID1 expression levels in WT, 20q gain, and hESC lines overexpressing BCL2L1, ID1, or both (BCL2L1/ID1). (C) Immunofluorescence staining of the VUB03 cell line: WT, 20q gain, BCL2L1-OE, ID1-OE, and double (BCL2L1 + ID1) overexpression lines. The final condition represents wild-type cells treated with SB and BMP4. Stained markers include PAX6 (yellow; neuroectoderm, panel 1), TFAP2A (magenta; surface ectoderm, panel 2), KRT8 (turquoise; surface ectoderm, panel 2), EPCAM (turquoise; surface ectoderm, panel 3), and OCT4 (magenta; pluripotency, panel 3). (D) Quantification of neuroectoderm immunostaining of BCL2L1-OE, ID1-OE, and double (BCL2L1 + ID1) overexpression lines showing marker intensity similar to . (E) Immunofluorescence staining of VUB03 cells after 14 days of spontaneous differentiation. Stained markers include PAX6 (yellow; neuroectoderm, panel 1), TFAP2A (magenta; surface ectoderm, panel 2), OCT4 (turquoise; pluripotency, panel 2), EPCAM (turquoise; surface ectoderm, panel 3), and KRT8 (magenta; surface ectoderm, panel 3). (F) Quantification of immunostaining of BCL2L1-OE, ID1-OE, and double (BCL2L1 + ID1) overexpression lines after spontaneous differentiation showing marker intensity similar to . (G) Fraction of marker-positive cells after 8 days of neuroectoderm differentiation. (H) Fraction of marker-positive cells after 14 days of spontaneous differentiation.

Article Snippet: The ID1 expression plasmid (TFORF2860) was obtained from Addgene (plasmid #143643), with ID1 expression driven by the EF-1α promoter.

Techniques: Over Expression, Quantitative RT-PCR, Expressing, Immunofluorescence, Staining, Immunostaining, Marker

Journal: Cancer Research Communications

Article Title: 3D “Emboli” Culture Models Epithelial Breast Cancer Cell Oxidative Mitochondrial Metabolism with Relevance for Lung Metastasis

doi: 10.1158/2767-9764.CRC-25-0587

Figure Lengend Snippet: EmC leads to induction of genes related to lung metastasis. A, qRT-PCR of mRNA levels for indicated genes in SUM149 and MDA-MB-468 cells under the indicated culture conditions ( n = 3). B, Differential expression of the indicated genes derived from the scRNA-seq data comparing EmC vs. SphC in SUM149 and IBC-3 cells as indicated. C, qRT-PCR (top, n = 3) of ID1 expression in SUM149 and MDA-MB-468 cultures as indicated. D, Western blot analysis of ID1 expression of cells as in C . Actin served as a loading control. E, Western blot analysis of ID1 and ID3 proteins in established SUM149 EmC cells that were treated with AGX51 at the indicated concentrations for 3 days. Actin served as a loading control. F, FC in viable SUM149 and MDA-MB-468 cells in established EmC following AGX51 treatment at the indicated concentrations for 3 days ( n = 3). G, FC in viable SUM149 and MDA-MB-468 cells in established EmC and SphC following AGX51 treatment (100 μmol/L) for 3 days ( n = 3). Quantitative data are shown as mean ± SEM, *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; n.s., not significant.

Article Snippet: Immunohistochemistry was performed according to standard procedures with primary antibodies for ID1 at 1:50 (Cell Signaling Technology #23369), Ki67 at 1:200 (Cell Signaling Technology #9027), or isotype control rabbit monoclonal IgG (Cell Signaling Technology # 3900).

Techniques: Quantitative RT-PCR, Quantitative Proteomics, Derivative Assay, Expressing, Western Blot, Control