rabbit anti tpcn2 monoclonal antibodies  (Alomone Labs)

Journal: Chinese Medical Journal

Article Title: Deficiency of two-pore segment channel 2 contributes to systemic lupus erythematosus via regulation of apoptosis and cell cycle

doi: 10.1097/CM9.0000000000001893

Figure Lengend Snippet: The primers used in the quantitative reverse transcription polymerase chain reaction.

Article Snippet: The membranes were blocked with 5% nonfat dry milk for 1 h, and then incubated with rabbit anti- TPCN2 monoclonal antibodies (1:200; Alomone Labs, Jerusalem, Israel) overnight at 4°C.

Techniques:

Journal: Chinese Medical Journal

Article Title: Deficiency of two-pore segment channel 2 contributes to systemic lupus erythematosus via regulation of apoptosis and cell cycle

doi: 10.1097/CM9.0000000000001893

Figure Lengend Snippet: TPCN2 -knockdown inhibits the proliferation in SLE. (A) Expression level of TPCN2 in PBMCs of SLE cases ( n = 6) and healthy controls ( n = 7). ∗ P < 0.05, Student's t test. Knockdown of TPCN2 with two independent shRNA in Jurkat (B) and THP-1 (C) cells; the left panel represents the relative expression levels of TPCN2 assessed by qRT-PCR; the right panel represents protein expression level of TPCN2 that was detected by Western blotting. The data represent the mean ( n = 3) ± SD. ∗ P < 0.05, Student's t test. Cell viability of silencing TPCN2 in Jurkat (D) and THP-1 (E) cells were determined by CCK-8 assay. The results represent the means ( n = 5) ± SD. Significant differences were evaluated using Student's t test. ∗ P < 0.05. CCK-8: Cell count kit-8; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; NC: Normal control; PBMCs: Peripheral blood mononuclear cells; qRT-PCR: Quantitative reverse transcription polymerase chain reaction; SD: Standard deviation; Sh#1: shTPCN2#1; Sh#2: shTPCN2#2; shRNA: Short hairpin RNA; SLE: Systemic lupus erythematosus; TPCN2 : Two-pore segment channel 2.

Article Snippet: The membranes were blocked with 5% nonfat dry milk for 1 h, and then incubated with rabbit anti- TPCN2 monoclonal antibodies (1:200; Alomone Labs, Jerusalem, Israel) overnight at 4°C.

Techniques: Expressing, shRNA, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Cell Counting, Reverse Transcription Polymerase Chain Reaction, Standard Deviation

Journal: Chinese Medical Journal

Article Title: Deficiency of two-pore segment channel 2 contributes to systemic lupus erythematosus via regulation of apoptosis and cell cycle

doi: 10.1097/CM9.0000000000001893

Figure Lengend Snippet: Knockdown of TPCN2 induces apoptosis and cell-cycle arrest in SLE. Apoptosis was detected by flow cytometry with Annexin V and PI in jurkat- TPCN2 -knockdown cells (A) and THP-1- TPCN2 -knockdown cells (B). TPCN2 -knockdown induced G2/M cell-cycle arrest. The percentage of G2-M phase cells of Jurkat (C) and THP-1 (D) was assessed by flow cytometry. ∗ P < 0.05. NC: Normal control; PI: Propidium iodide; Sh#1: shTPCN2#1; Sh#2: shTPCN2#2; SLE: Systemic lupus erythematosus; TPCN2 : Two-pore segment channel 2.

Article Snippet: The membranes were blocked with 5% nonfat dry milk for 1 h, and then incubated with rabbit anti- TPCN2 monoclonal antibodies (1:200; Alomone Labs, Jerusalem, Israel) overnight at 4°C.

Techniques: Flow Cytometry

Journal: Chinese Medical Journal

Article Title: Deficiency of two-pore segment channel 2 contributes to systemic lupus erythematosus via regulation of apoptosis and cell cycle

doi: 10.1097/CM9.0000000000001893

Figure Lengend Snippet: RNA-seq analyses of the effect of TPCN2 -knockdown on the gene expression profile. (A) The heatmap shows differential expression genes in sh#1 and sh#2 (in comparison with NC group). (B) The differential statistics of DEGs in NC groups vs . sh#1/sh#2. Blue columnar represent up-regulated DEGs. Red column represents down-regulated DEGs. (C) GO classification analysis of DEGs. NC group vs . sh#2. Molecular function – blue; Cellular components – red; Biological process – green. (D) Top 20 enriched KEGG pathways after silencing TPCN2 in Jurkat. The x-axis is the enrichment score, and the y-axis is the enriched pathways. DEGs: Differentially expressed genes; GO: Gene ontology; KEGG: Kyoto encyclopedia of genes and genomes; NC: Normal control; RNA-seq: RNA sequencing; Sh#1: shTPCN2#1; Sh#2: shTPCN2#2; TPCN2 : Two-pore segment channel 2.

Article Snippet: The membranes were blocked with 5% nonfat dry milk for 1 h, and then incubated with rabbit anti- TPCN2 monoclonal antibodies (1:200; Alomone Labs, Jerusalem, Israel) overnight at 4°C.

Techniques: RNA Sequencing Assay, Expressing

Journal: Chinese Medical Journal

Article Title: Deficiency of two-pore segment channel 2 contributes to systemic lupus erythematosus via regulation of apoptosis and cell cycle

doi: 10.1097/CM9.0000000000001893

Figure Lengend Snippet: Representative enriched pathways in high-risk shTPCN2#2 through GSEA analysis. GSEA results showed that the G2/M checkpoint (A), inflammatory response (B), complement (C), and PI3K-AKT-mTOR (D) pathways were enriched in the sh#2 expression group. Top panels indicate the enrichment scores for each gene. Bottom panels show the ranking metrics of each gene. Y-axis: ranking metric values; X-axis: ranks for all genes. (E) The mRNA expression of some DEGs in Jurkat cells were detected by qRT-PCR. The RNA was extracted from cells knocked down of TPCN2 with two independent shRNA. The results were shown as the mean ± SD from three independent experiments. ∗ P < 0.05. AKT: Protein kinase B; DEGs: Differentially expressed genes; GSEA: Gene set enrichment analysis; mTOR: Mechanistic target of rapamycin; NC: Normal control; NES: Normalized enrichment score; PI3K: Phosphatidylinositol 3-kinase; qRT-PCR: Quantitative reverse transcription polymerase chain reaction; SD: standard deviation; Sh#1: shTPCN2#1; Sh#2: shTPCN2#2; shRNA: Short hairpin RNA; TPCN2 : Two-pore segment channel 2.

Article Snippet: The membranes were blocked with 5% nonfat dry milk for 1 h, and then incubated with rabbit anti- TPCN2 monoclonal antibodies (1:200; Alomone Labs, Jerusalem, Israel) overnight at 4°C.

Techniques: Expressing, Quantitative RT-PCR, shRNA, Reverse Transcription Polymerase Chain Reaction, Standard Deviation