Journal: Scientific Reports

Article Title: Chemokines protect vascular smooth muscle cells from cell death induced by cyclic mechanical stretch

doi: 10.1038/s41598-017-15867-8

Figure Lengend Snippet: Expression of transcripts of candidate genes in RASMCs subjected to CMS. RASMCs were subjected to CMS for four hours, and expressions of transcripts of Cxcl1 ( a ), Cx3cl1 ( b ), Cxcl6 ( c ), Ccl12 ( d ), Nr4a1 ( e ), NOS2 ( f ), Mmp9 ( g ), Mmp13 ( h ), and Hspa1b ( i ) were evaluated using the real-time RT-PCR method. The quantity of the transcripts is expressed as a percentage of the control, normalized with respect to GAPDH. Data are means ± SE (n = 4); *p < 0.05 and **p < 0.01 versus control, and N.S. indicates no significant difference.

Article Snippet: ELISA kits for rat CXCL1 and CX3CL1 were purchased from RayBiotech, Inc. (Norcross, GA).

Techniques: Expressing, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: Chemokines protect vascular smooth muscle cells from cell death induced by cyclic mechanical stretch

doi: 10.1038/s41598-017-15867-8

Figure Lengend Snippet: Induction of CXCL1 and CX3CL1 in RASMCs subjected to CMS. RASMCs were incubated with SP600125 (20 μM) for 20 min and then subjected to CMS for four hours. Cells were harvested and analyzed by real-time RT-PCR with specific primers for Cxcl1 ( a ) or Cx3cl1 ( b ) or by immunoblotting using the anti-CX3CL1 antibody ( e ), and media were analyzed using ELISA for CXCL1 ( c ) or CX3CL1 ( d ). ( f ) The quantity of CX3CL1 expressed in RASMCs, as measured by scanning densitometry, is expressed as a percentage of the control, normalized with respect to beta-actin. Data are means ± SE (n = 3), and N.S. indicates no significant difference. RASMCs were incubated with BAY 11-7082 (5 µM) for 20 min and then subjected to CMS for four hours. Cells were harvested and analyzed by real-time RT-PCR with specific primers for Cxcl1 ( g ) or Cx3cl1 ( h ). The uncropped pictures of immuoblotting are shown in Supplemental Fig. ( a , b ).

Article Snippet: ELISA kits for rat CXCL1 and CX3CL1 were purchased from RayBiotech, Inc. (Norcross, GA).

Techniques: Incubation, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Chemokines protect vascular smooth muscle cells from cell death induced by cyclic mechanical stretch

doi: 10.1038/s41598-017-15867-8

Figure Lengend Snippet: Inhibition of CXCL1 and CX3CL1 accelerated cell death induced by CMS. RASMCs were incubated with SB265610 ( a – d ) or 18a ( e – h ) at the indicated concentrations and then incubated under normal conditions ( a , b , e , f ) or subjected to CMS for four hours ( c , d , g , h ). After CMS, the cells were incubated for 24 h, and cell viability and cell death were evaluated by the MTT assay ( a , c , e , g ) and the release of LDH ( b , d , f , h ), respectively. Cell viability and cell death are expressed as a percentage of the control. Data are means ± SE (n = 6); *p < 0.05 versus control, and N.S. indicates no significant difference; ANOVA, Tukey’s HSD test.

Article Snippet: ELISA kits for rat CXCL1 and CX3CL1 were purchased from RayBiotech, Inc. (Norcross, GA).

Techniques: Inhibition, Incubation, MTT Assay

Journal: Cell Division

Article Title: Anti-cancer effects of Coix seed extract through KCTD9-mediated ubiquitination of TOP2A in lung adenocarcinoma

doi: 10.1186/s13008-024-00112-2

Figure Lengend Snippet: The inhibiting effects of CSE on the immune escape of LUAD cells are reversed by KCTD9 knockdown. a Correlation between KCTD9 and T-cell infiltration in LUAD analyzed by TIMER 2.0 database. b PD-L1 mRNA expression in LUAD cells with different treatments detected by RT-qPCR assays. c PD-L1 protein expression in LUAD cells with different treatments detected by western blot assays. d - g The levels of TNF-α, IFN-γ, CXCL10, and CXCL9 in the co-culture system of LUAD cells and human CD8 + T cells. h Proliferation of human CD8 + T cells in co-culture system detected by CFSE assays. i The ability of differently treated LUAD cells against human CD8 + T cells was evaluated using tumor cell killing assay. Data are presented as the mean ± SD of results from at least three independent experiments performed in triplicates, two-way ANOVA with Tukey’s posttest, * p < 0.05

Article Snippet: The levels of tumor necrosis factor α (TNF-α), interferon-gamma (IFN-γ), CXCL10, and CXCL9 in the co-culture system were detected according to the instructions of Human TNF-α ELISA Kit (D711045, Shanghai Sangon Biological Engineering Technology & Services Co., Ltd., Shanghai, China), Human IFN-γ ELISA Kit (D711044, Sangon), Human CXCL10/IP-10 ELISA Kit (PC208, Beyotime), and MIG/CXCL9 Human ELISA Kit (EHCXCL9, Thermo Fisher).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Co-Culture Assay

Journal: Cell Division

Article Title: Anti-cancer effects of Coix seed extract through KCTD9-mediated ubiquitination of TOP2A in lung adenocarcinoma

doi: 10.1186/s13008-024-00112-2

Figure Lengend Snippet: Silencing of TOP2A reverses the effects of KCTD9 knockdown on LUAD cell immune escape. a Correlation of TOP2A expression in LUAD with human CD8 + T cell infiltration predicted in the TIMER database. b Correlation between TOP2A and PD-L1 expression predicted in the GEPIA database. c The prognostic outcomes in patients with high or low TOP2A expression are predicted in the Kaplan–Meier Plotter database. d The mRNA expression of TOP2A in LUAD cells after infection of sh-NC or sh-TOP2A was determined using RT-qPCR. e The mRNA expression of PD-L1 in LUAD cells was detected using RT-qPCR. f The protein expression of PD-L1 in LUAD cells was detected using western blot assays. g - j The levels of TNF-α, IFN-γ, CXCL10, and CXCL9 in the co-culture system of LUAD cells and human CD8 + T cells. k Proliferation of human CD8 + T cells in co-culture system detected by CFSE assays. l The ability of differently treated LUAD cells against human CD8 + T cells was evaluated using tumor cell killing assay. Data are presented as the mean ± SD of results from at least three independent experiments performed in triplicates, two-way ANOVA with Tukey’s posttest, * p < 0.05

Article Snippet: The levels of tumor necrosis factor α (TNF-α), interferon-gamma (IFN-γ), CXCL10, and CXCL9 in the co-culture system were detected according to the instructions of Human TNF-α ELISA Kit (D711045, Shanghai Sangon Biological Engineering Technology & Services Co., Ltd., Shanghai, China), Human IFN-γ ELISA Kit (D711044, Sangon), Human CXCL10/IP-10 ELISA Kit (PC208, Beyotime), and MIG/CXCL9 Human ELISA Kit (EHCXCL9, Thermo Fisher).

Techniques: Expressing, Infection, Quantitative RT-PCR, Western Blot, Co-Culture Assay

Journal: Cell Division

Article Title: Anti-cancer effects of Coix seed extract through KCTD9-mediated ubiquitination of TOP2A in lung adenocarcinoma

doi: 10.1186/s13008-024-00112-2

Figure Lengend Snippet: The inhibiting effects of CSE on the immune escape of LUAD cells in vivo are reversed by KCTD9 knockdown. a Tumor volume in mice in 3 weeks. b Weight changes in mouse tumors. c The protein expression of PD-L1 in mouse tumor tissues was examined using western blot assays. d Immunofluorescence analysis of CD8 + T cell infiltration in mouse tumor tissues. e The levels of CD8 + T cell activation markers GzmB and Perforin in the supernatant of the co-culture system were detected by ELISA. f Detection of apoptosis in mouse tumor tissues by TUNEL assay. The levels of TNF-α ( g ), IFN-γ ( h ), CXCL10 ( i ), and CXCL9 ( j ) in the mouse tumor tissues by ELISA. ( k ) The mRNA expression of KCTD9 in mouse tumor tissues was assessed by RT-qPCR. l The protein expression of TOP2A in mouse tumor tissues was assessed by western blot assays. Data are presented as the mean ± SD of results from at least three independent experiments performed in triplicates, one-way or two-way ANOVA with Tukey’s posttest, * p < 0.05

Article Snippet: The levels of tumor necrosis factor α (TNF-α), interferon-gamma (IFN-γ), CXCL10, and CXCL9 in the co-culture system were detected according to the instructions of Human TNF-α ELISA Kit (D711045, Shanghai Sangon Biological Engineering Technology & Services Co., Ltd., Shanghai, China), Human IFN-γ ELISA Kit (D711044, Sangon), Human CXCL10/IP-10 ELISA Kit (PC208, Beyotime), and MIG/CXCL9 Human ELISA Kit (EHCXCL9, Thermo Fisher).

Techniques: In Vivo, Expressing, Western Blot, Immunofluorescence, Activation Assay, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, TUNEL Assay, Quantitative RT-PCR