Journal: Scientific Reports

Article Title: Inactivation of TRPM7 kinase in mice results in enlarged spleens, reduced T-cell proliferation and diminished store-operated calcium entry

doi: 10.1038/s41598-018-21004-w

Figure Lengend Snippet: TRPM7/TRPM6 protein expression and TRPM7 kinase activity in splenic T cells. ( A ) Western blot analysis of immunoprecipitated TRPM7 from whole cell lysates of WT and KD splenic T cells. T cells were stimulated with PMA/ionomycin or anti-CD3/CD28 antibody coated beads for 48 hrs. Mouse embryonic fibroblasts were used as a positive control. Equal amounts of protein before immunoprecipitation were ensured by probing for actin. ( B ) Incorporation of 32 P into exogenous myelin basic protein (MBP) by TRPM7 immunoprecipitated from WT and KD resting T cells. Equal quantities of MBP were verified by coomassie blue staining. ( C ) Control experiment showing that anti-TRPM6 antibody was able to recognize TRPM6, by immunoprecipitation using anti-TRPM6 antibody in GFP-TRPM6 transfected HEK cells ( D ). Western blot analysis of TRPM6 immunoprecipitated from WT and KD mouse T cells and kidneys. Full gel images are provided in Supplementary Fig. .

Article Snippet: Extracts of T cells, mouse embryonic fibroblasts (MEF) or HEK293 cells (untransfected or heterologously expressing GFP tagged human TRPM6 in pEGFP-C1 vector) were obtained as described above for the TRPM7 kinase assay and incubated with anti-TRPM7 antibody (1:500) or anti-TRPM6 antibody (1:500, ACC-046; Alomone labs, Israel) overnight at 4 °C, followed by incubation with protein A sepharose beads for 1 hr.

Techniques: Expressing, Activity Assay, Western Blot, Immunoprecipitation, Positive Control, Staining, Transfection

Journal: eLife

Article Title: SAFB regulates hippocampal stem cell fate by targeting Drosha to destabilize Nfib mRNA

doi: 10.7554/eLife.74940

Figure Lengend Snippet:

Article Snippet: Recombinant DNA reagent , pEGFPC1-6XHis- FLKSRP (plasmid) , , RRID: Addgene_23001 , pEGFPC1 expression vector.

Techniques: Control, Protease Inhibitor, Recombinant, Cloning, Bicinchoninic Acid Protein Assay, Mutagenesis, Transfection, Sequencing, esiRNA, Blocking Assay, Plasmid Preparation, Expressing, Software

Journal: Frontiers in Endocrinology

Article Title: Analysis of the key ligand receptor CADM1_CADM1 in the regulation of thyroid cancer based on scRNA-seq and bulk RNA-seq data

doi: 10.3389/fendo.2022.969914

Figure Lengend Snippet: Screening of crucial L/R pairs. (A) Venn diagram of 38 L/R pairs communicating between malignant cells/non-malignant cells and 44 L/R pairs communicating between non-malignant cells/malignant cells. (B) Forest plot of univariate Cox regression analysis of 16 L/R pairs. (C) KM curves of CADM1-CADM1.

Article Snippet: The eukaryotic expression vector pEGFP-C1-CADM1 was purchased from Genepharma (Shanghai, China).

Techniques:

Journal: Frontiers in Endocrinology

Article Title: Analysis of the key ligand receptor CADM1_CADM1 in the regulation of thyroid cancer based on scRNA-seq and bulk RNA-seq data

doi: 10.3389/fendo.2022.969914

Figure Lengend Snippet: Differential analysis of 16 L/R pairs.

Article Snippet: The eukaryotic expression vector pEGFP-C1-CADM1 was purchased from Genepharma (Shanghai, China).

Techniques:

Journal: Frontiers in Endocrinology

Article Title: Analysis of the key ligand receptor CADM1_CADM1 in the regulation of thyroid cancer based on scRNA-seq and bulk RNA-seq data

doi: 10.3389/fendo.2022.969914

Figure Lengend Snippet: Differences in the expression of CADM1-CADM1 in clinical phenotypes (** P < 0.01, *P < 0.05). The differences in CADM1-CADM1 by gender (A) , T stage (B) , M stage (C) , N stage (D) , stage (E) and age (F) . ns, no significance.

Article Snippet: The eukaryotic expression vector pEGFP-C1-CADM1 was purchased from Genepharma (Shanghai, China).

Techniques: Expressing

Journal: Frontiers in Endocrinology

Article Title: Analysis of the key ligand receptor CADM1_CADM1 in the regulation of thyroid cancer based on scRNA-seq and bulk RNA-seq data

doi: 10.3389/fendo.2022.969914

Figure Lengend Snippet: Analysis of differential genes associated with CADM1-CADM1. (A) Volcano map showing differential genes between tumor tissues and para-tumors. (B) Bidirectional Clustering Heatmap showing the relationship between CADM1-CADM1 and differential genes associated with CADM1-CADM1. Each column in the heatmap represents a sample, and each row represents the expression level of a gene. The color scale beside the heatmap represents the raw Z-score: blue (low expression), red (high expression). (C) Scatter plot of 9 enriched GO terms of molecular function (MF). (D) Scatter plot of top 10 enriched GO terms of biological process (BP). (E) Scatter plot of top 10 enriched GO terms of cellular component (CC). (F) KEGG enrichment analysis of differential genes associated with CADM1-CADM1.

Article Snippet: The eukaryotic expression vector pEGFP-C1-CADM1 was purchased from Genepharma (Shanghai, China).

Techniques: Expressing

Journal: Frontiers in Endocrinology

Article Title: Analysis of the key ligand receptor CADM1_CADM1 in the regulation of thyroid cancer based on scRNA-seq and bulk RNA-seq data

doi: 10.3389/fendo.2022.969914

Figure Lengend Snippet: Information of pathways involved in CADM1-CADM1.

Article Snippet: The eukaryotic expression vector pEGFP-C1-CADM1 was purchased from Genepharma (Shanghai, China).

Techniques:

Journal: Frontiers in Endocrinology

Article Title: Analysis of the key ligand receptor CADM1_CADM1 in the regulation of thyroid cancer based on scRNA-seq and bulk RNA-seq data

doi: 10.3389/fendo.2022.969914

Figure Lengend Snippet: Box plots showing ssGSEA scores of related signaling pathways in high and low expression groups of CADM1-CADM1 (****P < 0.0001, *P < 0.05). (A) PI3K-Akt signaling pathway. (B) MAPK signaling pathway. (C) Wnt signaling pathway. (D) TGF-β signaling pathway. (E) JAK-STAT signaling pathway. (F) cAMP signaling pathway.

Article Snippet: The eukaryotic expression vector pEGFP-C1-CADM1 was purchased from Genepharma (Shanghai, China).

Techniques: Expressing

Journal: Frontiers in Endocrinology

Article Title: Analysis of the key ligand receptor CADM1_CADM1 in the regulation of thyroid cancer based on scRNA-seq and bulk RNA-seq data

doi: 10.3389/fendo.2022.969914

Figure Lengend Snippet: The relationship between CADM1-CADM1 and DNA, RNA methylation. (A) Comparison of CADM1 methylation in high CADM1-CADM1 group and low CADM1-CADM1 group. (B) Correlation analysis of CADM1-CADM1 and CADM1 methylation. (C–E) Comparison of the scores of erasers, readers, writers in m1a, m6a, m5c modification between high CADM1-CADM1 group and low CADM1-CADM1 group separately (****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05).

Article Snippet: The eukaryotic expression vector pEGFP-C1-CADM1 was purchased from Genepharma (Shanghai, China).

Techniques: Methylation, Modification

Journal: Frontiers in Endocrinology

Article Title: Analysis of the key ligand receptor CADM1_CADM1 in the regulation of thyroid cancer based on scRNA-seq and bulk RNA-seq data

doi: 10.3389/fendo.2022.969914

Figure Lengend Snippet: The relationship between CADM1-CADM1, gender, age, stage, T stage, M stage, N stage and genes associated with apoptosis, ferroptosis, pyroptosis and cellular senescence.

Article Snippet: The eukaryotic expression vector pEGFP-C1-CADM1 was purchased from Genepharma (Shanghai, China).

Techniques:

Journal: Frontiers in Endocrinology

Article Title: Analysis of the key ligand receptor CADM1_CADM1 in the regulation of thyroid cancer based on scRNA-seq and bulk RNA-seq data

doi: 10.3389/fendo.2022.969914

Figure Lengend Snippet: Comparison of targeted drug sensitivity in high CADM1-CADM1 group and low CADM1-CADM1 group. (****P < 0.0001, ***P < 0.001, *P < 0.05). (A) Comparison of the sensitivity of Erlotinib. (B) Comparison of the sensitivity of MG-132. (C) Comparison of the sensitivity of AZ628. (D) Comparison of the sensitivity of Sorafenib. (E) Comparison of the sensitivity of Saracatinib. (F) Comparison of the sensitivity of Dasatinib. (G) Comparison of the sensitivity of Parthenolide. (H) Comparison of the sensitivity of Bortezomib. (I) Comparison of the sensitivity of Shikonin.

Article Snippet: The eukaryotic expression vector pEGFP-C1-CADM1 was purchased from Genepharma (Shanghai, China).

Techniques:

Journal: Frontiers in Endocrinology

Article Title: Analysis of the key ligand receptor CADM1_CADM1 in the regulation of thyroid cancer based on scRNA-seq and bulk RNA-seq data

doi: 10.3389/fendo.2022.969914

Figure Lengend Snippet: The differential expression of CADM1-CADM1 between tumor tissues and normal tissues in pan-cancer (****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05).

Article Snippet: The eukaryotic expression vector pEGFP-C1-CADM1 was purchased from Genepharma (Shanghai, China).

Techniques: Expressing

Journal: Frontiers in Endocrinology

Article Title: Analysis of the key ligand receptor CADM1_CADM1 in the regulation of thyroid cancer based on scRNA-seq and bulk RNA-seq data

doi: 10.3389/fendo.2022.969914

Figure Lengend Snippet: The expression of CADM1-CADM1 in PTC cell lines. (A) The different expression of CADM1 in N3, K1, BAPAP, TPC1 and IHH4 cell lines. (B) Results of immunofluorescence staining of IHH4 cells, which were mainly located in cytoplasm and a little in nucleus. (C) Results of immunofluorescence staining of TPC1 cells, which were also mainly located in cytoplasm and a little in nucleus. (D–I) Overexpression of CADM1 in IHH4 cell lines by pEGFP-C1-CADM1 transfection inhibits cell migration. D, Blank control (IHH4) (×200). E, pEGFP-C1 (NC) (×200). F, pEGFP-C1-CADM1 (CADM1) (×200). (G–I) Overexpression of CADM1 in TPC1 cell lines by pEGFP-C1-CADM1 transfection inhibits cell migration. G, Blank control (TPC1) (×200).H, pEGFP-C1 (NC) (×200). I, pEGFP-C1-CADM1 (CADM1) (×200).

Article Snippet: The eukaryotic expression vector pEGFP-C1-CADM1 was purchased from Genepharma (Shanghai, China).

Techniques: Expressing, Immunofluorescence, Staining, Over Expression, Transfection, Migration

Journal: bioRxiv

Article Title: Endosomal Chemokine Receptor Signalosomes Regulate Central Mechanisms Underlying Cell Migration

doi: 10.1101/2022.09.27.509755

Figure Lengend Snippet: Endosomal G i/o activation by internalized CCR7. (A) Confocal microscopy imaging displaying HEK293-CCR7 cells transiently expressing the early endosomal marker Rab5a-RFP. The cells were treated with either 100 nM CCL19-Alexa488, 100 nM CCL21-Alexa488, or Ni-NTA-Alexa488 for 15 min followed by extensive washout to remove extracellular chemokines. The cells were imaged 15-30 min after this washout step. (B) Schematic representation of the EbBRET-based assay to monitor proximity between RlucII-miniGi and the endosomal marker rGFP-Rab5 upon CCR7 activation from endosomes. (C) EbBRET measurements from CCR7-expressing HEK293 cells co-transfected with RlucII-miniGi and rGFP-Rab5 upon stimulation with 100 nM CCL19, 100 nM CCL21, or vehicle control. Data represents the mean ±SE from N=3 independent experiments. One-way ANOVA with Turkey’s multiple comparison post hoc test was applied to determine statistical differences between the measurements (** p < 0.01; *** p < 0.001). (D) Confocal microscopy imaging displaying CCR7-expressing HEK293 cells cotransfected with the plasma membrane marker RFP-Lck and Halo-miniGi. The cells were stimulated with 100 nM CCL19, 100 nM CCL21, or vehicle control for 10 min. (E) Confocal microscopy imaging displaying CCR7-expressing HEK293 cells co-transfected with the endosomal marker RFP-EEA1 and Halo-miniGi. The cells were stimulated with 100 nM CCL19, 100 nM CCL21, or vehicle control for 30 min.

Article Snippet: RFP-Lck (C-tRFP-Lck cloned into PCMV6-AC-RFP expression vector) and RFP-EEA1 (TagRFP-T-EEA1 cloned into pEGFP-C1 vector) were purchased from Addgene (respectively #RC100049 and #42635).

Techniques: Activation Assay, Confocal Microscopy, Imaging, Expressing, Marker, Transfection