Journal: Journal of Medical Virology

Article Title: SARS‐CoV‐2 pseudovirus infectivity and expression of viral entry‐related factors ACE2, TMPRSS2, Kim‐1, and NRP‐1 in human cells from the respiratory, urinary, digestive, reproductive, and immune systems

doi: 10.1002/jmv.27244

Figure Lengend Snippet: Cell lines used in the present study

Article Snippet: 769‐P , Renal cell adenocarcinoma , ATCC CRL‐1933 , RPMI‐1640 complete medium.

Techniques: Cell Culture

Journal: Cancer Science

Article Title: LINC00973 is involved in cancer immune suppression through positive regulation of Siglec‐15 in clear‐cell renal cell carcinoma

doi: 10.1111/cas.14611

Figure Lengend Snippet: High expression of LINC00973 in clear‐cell renal cell carcinoma (ccRCC). A, Real‐time PCR analysis of relative expression of LINC00973 in ccRCC tumor tissues and paired adjacent normal tissues. B, Kaplan‐Meier survival curve of ccRCC patients with either high or low level of LINC00973. C, Correlation analysis of relative LINC00973 level with tumor stage of ccRCC patients (KIRC) in the publicly available TCGA database. D, Overall survival analysis of KIRC dataset in respect to LINC00973 levels with Gepia ( http://gepia.cancer‐pku.cn/ ). E, Quantitative PCR analysis of relative expression of LINC00973 in panel of ccRCC cell lines including 769‐p, Caki‐1, Caki‐2, A498, A704, ACHN, RCC4, and 786‐0. F, Subcellular localization of LINC00973 was determined by fractionization analysis with GAPDH as a cytosol marker and U6 as a nucleus marker, respectively

Article Snippet: Eight human ccRCC cell lines (786‐0, 769‐p, A704, A498, ACNH, Caki‐1, Caki‐2, and RCC4) and an immortalized kidney proximal tubule epithelial cell line HK‐2 were obtained from the ATCC.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Marker

Journal: Cancer Science

Article Title: LINC00973 is involved in cancer immune suppression through positive regulation of Siglec‐15 in clear‐cell renal cell carcinoma

doi: 10.1111/cas.14611

Figure Lengend Snippet: LINC00973 upregulated Siglec‐15 in clear‐cell renal cell carcinoma (ccRCC) cells. (A) Comparison of LINC00973 abundance in Siglec‐15 negative and positive ccRCC tumors. (B) Representative images of Siglec‐15 immunohistochemistry (IHC) staining results. (C) Correlation analysis of endogenous LINC00973 and Siglec‐15 expression in publicly available TCGA kidney cells (n = 34). (D) Correlation analysis of LINC00973 and Siglec‐15 in clinical ccRCC tumor samples (n = 100). (E) Establishment of LINC00973‐knockdown cell lines in 769‐p and Caki‐1 cells was validated by real‐time PCR. (F) Establishment of LINC00973‐overexpressing cell lines in A704 and A498 cells was validated by real‐time PCR. (G) Relative expression of Siglec‐15 transcripts in LINC00973‐deficient 769‐p and Caki‐1 cells. (H). Relative expression of Siglec‐15 transcripts in LINC00973‐proficient A704 and A498 cells. (I) Western blot analysis of Siglec‐15 protein in LINC00973 knockdown or overexpressing cells. Flow cytometry analysis of cell surface Siglec‐15 abundance in response to LINC00973 knockdown in 769‐p (J) and Caki‐1 (K) cells, and LINC00973‐overexpressing in A704 (L) and A498 (M). N. Immunofluorescence staining of cell surface Siglec‐15 of A498 in response to either control (NC) or LINC00973 overexpression

Article Snippet: Eight human ccRCC cell lines (786‐0, 769‐p, A704, A498, ACNH, Caki‐1, Caki‐2, and RCC4) and an immortalized kidney proximal tubule epithelial cell line HK‐2 were obtained from the ATCC.

Techniques: Comparison, Immunohistochemistry, Expressing, Knockdown, Real-time Polymerase Chain Reaction, Western Blot, Flow Cytometry, Immunofluorescence, Staining, Control, Over Expression

Journal: Cancer Science

Article Title: LINC00973 is involved in cancer immune suppression through positive regulation of Siglec‐15 in clear‐cell renal cell carcinoma

doi: 10.1111/cas.14611

Figure Lengend Snippet: Siglec‐15 was a direct target of miR‐7109 in clear‐cell renal cell carcinoma (ccRCC). (A) Real‐time PCR analysis of miR‐7109 expression in ccRCC tumor samples with paired adjacent normal tissues. (B) Correlation analysis between miR‐7109 and Siglec‐15 in ccRCC tissue samples (n = 100). (C) Change of miR‐7109 expression in both A704 and A498 cells in response to miR‐7109 inhibitor. (D) Change of miR‐7109 expression in both 769‐p and Caki‐1 cells in response to miR‐7109 mimic. (E) Endogenous Siglec‐15 transcripts were quantified in both A704 and A498 cells transfected with either control or miR‐7109 inhibitor. (F) Endogenous Siglec‐15 transcripts were quantified in both 769‐p and Caki‐1 cells transfected with either control or miR‐7109 mimics. (G) Western blot analysis of Siglec‐15 proteins in the same cells as in panel E and F. (H) Flow cytometry analysis of cell surface Siglec‐15 abundance in A704 and A498 cells in response to miR‐7109 inhibitor. (I) Flow cytometry analysis of cell surface Siglec‐15 abundance in 769‐p and Caki‐1 cells in response to miR‐7109 mimics. (J) Immunofluorescence staining of cell surface Siglec‐15 in A498 cells transfected with either control or miR‐7109 inhibitor. (K) Alignment between miR‐7109 seed region and both wild‐type and putative binding site‐mutated Siglec‐15 3ʹUTR. (L) Luciferase reporter analysis of the potential regulatory effects of miR‐7109 on Siglec‐15 in 293T cells, which were transfected with control, miR‐7109 mimics, and miR‐7109 inhibitors, respectively

Article Snippet: Eight human ccRCC cell lines (786‐0, 769‐p, A704, A498, ACNH, Caki‐1, Caki‐2, and RCC4) and an immortalized kidney proximal tubule epithelial cell line HK‐2 were obtained from the ATCC.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Transfection, Control, Western Blot, Flow Cytometry, Immunofluorescence, Staining, Binding Assay, Luciferase

Journal: Cancer Science

Article Title: LINC00973 is involved in cancer immune suppression through positive regulation of Siglec‐15 in clear‐cell renal cell carcinoma

doi: 10.1111/cas.14611

Figure Lengend Snippet: LINC00973 functioned as ceRNA against miR‐7109 in clear‐cell renal cell carcinoma (ccRCC). (A) Relative expression of miR‐7109 in both 769‐p and Caki‐1 cells transfected with either negative control or LINC00973‐specific shRNA. (B) Relative expression of miR‐7109 in both A704 and A498 cells transfected with either control or LINC00973‐expressing plasmids. (C) RNA‐pulldown results in both A704 and A498 cells with control, wild‐type LINC0973 fragment, or corresponding mutant, respectively. (D) Relative expression of LINC00973 in both A704 and A498 cells transfected with either negative control or miR‐7109 inhibitors. (E) Relative expression of LINC00973 in both 769‐p and Caki‐1 cells transfected with either control or miR‐7109 mimics. (F) RNA‐pulldown results in both 769‐p and Caki‐1 cells with either control or miR‐7109. (G) Reverse correlation analysis between endogenous miR‐7109 and LINC00973 in ccRCC tumor samples (n = 100). (H) Illustration of aligned miR‐7109 with both wild‐type and mutated LINC00973. (I) Luciferase reporter results in 293T cells co–transfected with LINC00973 (wt and mut) and miR‐NC, miR‐7109 mimic, and miR‐7109 inhibitor, respectively

Article Snippet: Eight human ccRCC cell lines (786‐0, 769‐p, A704, A498, ACNH, Caki‐1, Caki‐2, and RCC4) and an immortalized kidney proximal tubule epithelial cell line HK‐2 were obtained from the ATCC.

Techniques: Expressing, Transfection, Negative Control, shRNA, Control, Mutagenesis, Luciferase

Journal: Cancer Science

Article Title: LINC00973 is involved in cancer immune suppression through positive regulation of Siglec‐15 in clear‐cell renal cell carcinoma

doi: 10.1111/cas.14611

Figure Lengend Snippet: LINC00973‐miR‐7109‐Siglec‐15 axis involved in immune activations in clear‐cell renal cell carcinoma (ccRCC) cells. (A) Establishment of Siglec‐15 overexpression cells in A704 and A498 was confirmed by real‐time PCR. (B) Establishment of Siglec‐15 knockdown cells in 769‐p and Caki‐1 cells was confirmed by real‐time PCR. (C) Western blot results of relative expression of Siglec‐15 protein in cells described in (A) and (B). (D) Interleukin‐2 (IL‐2) production analysis in Jurkat: A704/A498 (EV or Siglec‐15, 2:1) co–culture system. IL‐2 was quantified with an ELISA kit at 48 and 72 h, respectively. (E) IL‐2 production analysis in Jurkat: 769‐p/Caki‐1 (shNT or shSiglec‐15, 2:1) co–culture system. (F) IL‐2 production analysis of Jurkat co–culture system with both A704 and A498 cells transfected with control or LINC00973 in combination with either miR‐NC or miR7109 mimics. (G) IL‐2 production analysis of Jurkat co–culture system with both 769‐p and Caki‐1 cells transfected with negative control or shLINC0973 in combination with either miR‐NC or miR‐7109 inhibitor. (H) Tumor necrosis factor‐α (TNF‐α) secretion analysis of Jurkat co–culture system with both A704 and A498 cells transfected with control or LINC00973 in combination with either miR‐NC or miR‐7109 mimics. (I) TNF‐α secretion analysis of Jurkat co–culture system with both 769‐p and Caki‐1 cells transfected with either negative control or shLINC00973 in combination with either miR‐NC or miR‐7109 inhibitors

Article Snippet: Eight human ccRCC cell lines (786‐0, 769‐p, A704, A498, ACNH, Caki‐1, Caki‐2, and RCC4) and an immortalized kidney proximal tubule epithelial cell line HK‐2 were obtained from the ATCC.

Techniques: Over Expression, Real-time Polymerase Chain Reaction, Knockdown, Western Blot, Expressing, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Transfection, Control, Negative Control