Journal: European Journal of Histochemistry : EJH

Article Title: SNORA38b promotes proliferation, migration, invasion and epithelial-mesenchymal transition of gallbladder cancer cells via activating TGF-β/Smad2/3 signaling

doi: 10.4081/ejh.2023.3899

Figure Lengend Snippet: Primer sequences.

Article Snippet: Meanwhile, siRNA NC and SNORA38B siRNAs (SNORA38B siRNA1, SNORA38B siRNA2 and SNORA38B siRNA3; RiboBio Co. Ltd., Guangzhou, China) were transfected into GBC cells using the Lipofectamine 2000 reagent to downregulate SNORA38B level in GBC cells.

Techniques:

Journal: European Journal of Histochemistry : EJH

Article Title: SNORA38b promotes proliferation, migration, invasion and epithelial-mesenchymal transition of gallbladder cancer cells via activating TGF-β/Smad2/3 signaling

doi: 10.4081/ejh.2023.3899

Figure Lengend Snippet: SNORA38B was remarkably elevated in GBC tissues. A ) The level of SNORA38B in gallbladder cancer tissues and normal tissues was evaluated using RT-qPCR. B-E) RT-qPCR analysis of SNORA38B level in GBC-SD and NOZ cells transfected with ( B,C ) SNORA38B-OE or ( D, E ) si-SNORA38B-1, -2, or -3. F ) GBC-SD and NOZ cells were transfected with SNORA38B-OE or si- SNORA38B-2. Cell viability was assessed using CCK-8 assay. **p<0.01.

Article Snippet: Meanwhile, siRNA NC and SNORA38B siRNAs (SNORA38B siRNA1, SNORA38B siRNA2 and SNORA38B siRNA3; RiboBio Co. Ltd., Guangzhou, China) were transfected into GBC cells using the Lipofectamine 2000 reagent to downregulate SNORA38B level in GBC cells.

Techniques: Quantitative RT-PCR, Transfection, CCK-8 Assay

Journal: European Journal of Histochemistry : EJH

Article Title: SNORA38b promotes proliferation, migration, invasion and epithelial-mesenchymal transition of gallbladder cancer cells via activating TGF-β/Smad2/3 signaling

doi: 10.4081/ejh.2023.3899

Figure Lengend Snippet: SNORA38B deficiency suppressed GBC cell proliferation, migration and invasion. GBC-SD cells were transfected with SNORA38B-OE or si-SNORA38B-2. A ) Cell proliferation was assessed using EdU staining assay. B ) Cell migration and invasion were detected by transwell assays. **p<0.01.

Article Snippet: Meanwhile, siRNA NC and SNORA38B siRNAs (SNORA38B siRNA1, SNORA38B siRNA2 and SNORA38B siRNA3; RiboBio Co. Ltd., Guangzhou, China) were transfected into GBC cells using the Lipofectamine 2000 reagent to downregulate SNORA38B level in GBC cells.

Techniques: Migration, Transfection, Staining

Journal: European Journal of Histochemistry : EJH

Article Title: SNORA38b promotes proliferation, migration, invasion and epithelial-mesenchymal transition of gallbladder cancer cells via activating TGF-β/Smad2/3 signaling

doi: 10.4081/ejh.2023.3899

Figure Lengend Snippet: SNORA38B deficiency induced GBC cell apoptosis. GBC-SD cells were transfected with SNORA38B-OE or si-SNORA38B- 2. A,B ) Cell apoptosis was evaluated using ( A ) flow cytometry and ( B ) TUNEL staining assays. C ) IF staining assay was performed to assess caspase 3 expression in cells. **p<0.01.

Article Snippet: Meanwhile, siRNA NC and SNORA38B siRNAs (SNORA38B siRNA1, SNORA38B siRNA2 and SNORA38B siRNA3; RiboBio Co. Ltd., Guangzhou, China) were transfected into GBC cells using the Lipofectamine 2000 reagent to downregulate SNORA38B level in GBC cells.

Techniques: Transfection, Flow Cytometry, TUNEL Assay, Staining, Expressing

Journal: European Journal of Histochemistry : EJH

Article Title: SNORA38b promotes proliferation, migration, invasion and epithelial-mesenchymal transition of gallbladder cancer cells via activating TGF-β/Smad2/3 signaling

doi: 10.4081/ejh.2023.3899

Figure Lengend Snippet: SNORA38B deficiency suppressed the EMT of GBC cells. GBC-SD cells were transfected with SNORA38B-OE or si- SNORA38B-2. A-C ) IF staining assay was applied to evaluate E-cadherin, N-cadherin, and vimentin expressions in cells. **p<0.01.

Article Snippet: Meanwhile, siRNA NC and SNORA38B siRNAs (SNORA38B siRNA1, SNORA38B siRNA2 and SNORA38B siRNA3; RiboBio Co. Ltd., Guangzhou, China) were transfected into GBC cells using the Lipofectamine 2000 reagent to downregulate SNORA38B level in GBC cells.

Techniques: Transfection, Staining

Journal: European Journal of Histochemistry : EJH

Article Title: SNORA38b promotes proliferation, migration, invasion and epithelial-mesenchymal transition of gallbladder cancer cells via activating TGF-β/Smad2/3 signaling

doi: 10.4081/ejh.2023.3899

Figure Lengend Snippet: SNORA38B deficiency suppressed GBC cell growth via TGF-β signaling. GBC-SD cells were treated with si-SNORA38B-2, TGF-β1 or TGF-β1 + si-SNORA38B-2. A ) Cell viability was assessed using CCK-8 assay. B ) Cell proliferation was assessed using EdU staining assay. C ) Cell apoptosis was evaluated using TUNEL staining assay. D ) Cell migration and invasion were detected by transwell assays. **p<0.01.

Article Snippet: Meanwhile, siRNA NC and SNORA38B siRNAs (SNORA38B siRNA1, SNORA38B siRNA2 and SNORA38B siRNA3; RiboBio Co. Ltd., Guangzhou, China) were transfected into GBC cells using the Lipofectamine 2000 reagent to downregulate SNORA38B level in GBC cells.

Techniques: CCK-8 Assay, Staining, TUNEL Assay, Migration

Journal: European Journal of Histochemistry : EJH

Article Title: SNORA38b promotes proliferation, migration, invasion and epithelial-mesenchymal transition of gallbladder cancer cells via activating TGF-β/Smad2/3 signaling

doi: 10.4081/ejh.2023.3899

Figure Lengend Snippet: SNORA38B deficiency suppressed the EMT of GBC cells via TGF-β signaling. GBC-SD cells were treated with si- SNORA38B-2, TGF-β1 or TGF-β1 + si-SNORA38B-2. A-C ) IF staining assay was applied for evaluating E-cadherin, N-cadherin, and vimentin expressions in cells. **p<0.01.

Article Snippet: Meanwhile, siRNA NC and SNORA38B siRNAs (SNORA38B siRNA1, SNORA38B siRNA2 and SNORA38B siRNA3; RiboBio Co. Ltd., Guangzhou, China) were transfected into GBC cells using the Lipofectamine 2000 reagent to downregulate SNORA38B level in GBC cells.

Techniques: Staining

Journal: European Journal of Histochemistry : EJH

Article Title: SNORA38b promotes proliferation, migration, invasion and epithelial-mesenchymal transition of gallbladder cancer cells via activating TGF-β/Smad2/3 signaling

doi: 10.4081/ejh.2023.3899

Figure Lengend Snippet: SNORA38B deficiency suppressed the growth of GBC cells via inactivating TGF-β/Smad2/3 signaling. GBC-SD cells were treated with si-SNORA38B-2, TGF-β1 or TGF-β1 + si-SNORA38B-2. A,B ) Western blot assay was applied for detecting p-Smad2 and p-Smad3 levels in cells. **p<0.01.

Article Snippet: Meanwhile, siRNA NC and SNORA38B siRNAs (SNORA38B siRNA1, SNORA38B siRNA2 and SNORA38B siRNA3; RiboBio Co. Ltd., Guangzhou, China) were transfected into GBC cells using the Lipofectamine 2000 reagent to downregulate SNORA38B level in GBC cells.

Techniques: Western Blot

Journal: European Journal of Histochemistry : EJH

Article Title: SNORA38b promotes proliferation, migration, invasion and epithelial-mesenchymal transition of gallbladder cancer cells via activating TGF-β/Smad2/3 signaling

doi: 10.4081/ejh.2023.3899

Figure Lengend Snippet: SNORA38B deficiency triggered ferroptosis of GBC cells via TGF-β signaling. GBC-SD cells were treated with si- SNORA38B-2, TGF-β1 or TGF-β1 + si-SNORA38B-2. A,B ) The levels of ROS and Fe 2+ levels in cells were detected using ELISA. **p<0.01.

Article Snippet: Meanwhile, siRNA NC and SNORA38B siRNAs (SNORA38B siRNA1, SNORA38B siRNA2 and SNORA38B siRNA3; RiboBio Co. Ltd., Guangzhou, China) were transfected into GBC cells using the Lipofectamine 2000 reagent to downregulate SNORA38B level in GBC cells.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Viruses

Article Title: Effect of the Interaction between Viral PB2 and Host SphK1 on H9N2 AIV Replication in Mammals

doi: 10.3390/v14071585

Figure Lengend Snippet: Transcriptomic screening for key differentially expressed host SphK1 gene. ( A ) The number of differential genes in each group was obtained by transcriptomic detection. Blue and gray represent up-regulated and down-regulated differential genes, respectively, and the numbers on the columns represent the number of differential genes; ( B ) Venn diagram of the number of differential genes compared between different groups, with different colors indicating different combinations of comparisons; ( C ) fold change value of SphK1 gene after influenza virus infection compared with the Mock group, presented as a heat map, the darker the red color, the larger the value; ( D ) schematic diagram of the role of SphK1 in cellular metabolism; ( E ) A549 cells were infected with the virus at MOI = 0.2, Cells were collected 24 h after infection to detect the mRNA expression of SphK1; ( F ) A549 cells were infected with the virus at MOI = 0.2, and the cells were collected 24 h after infection to detect the expression of SphK1 protein, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a loading control. ( G ) for ( F ) densitometry analysis quantification. Differences were considered statistically significant at p < 0.05 (** p < 0.01, ANOVA).

Article Snippet: siRNAs against SphK1 were transfected using Lipofectamine RNAiMax (Invitrogen, Carlsbad, CA, USA) at a final concentration of 20 nM following the manufacturer’s instructions. siRNAs against SphK1 (siRNA1: 5′-GCAGCUUCCUUGAACCAUUTT-3′, siRNA2: 5′-GAUUGCUGAUGUGGACCUTAA-3′, siRNA3: 5′-GUGCACCCAAACUACUUCUTT-3′) are constructed by GenePharma Corporation (Shanghai, China).

Techniques: Infection, Expressing

Journal: Viruses

Article Title: Effect of the Interaction between Viral PB2 and Host SphK1 on H9N2 AIV Replication in Mammals

doi: 10.3390/v14071585

Figure Lengend Snippet: SphK1 protein inhibits H9N2 virus protein expression and virus replication in A549 cells. ( A ) Three designed siRNAs targeting SphK1 and negative control (NC) were transfected into A549 cells, and cells were collected 24 h after transfection for western blotting (WB) detection and ( B ) densitometry analysis and quantification; ( C ) siRNA 1 and NC was pretransfected into A549 cells, the virus was then infected with the virus at MOI = 0.5, and the cells were collected for WB at 4, 8, 12, 24 and 36 h after infection, and the expression levels of viral NP, M1 and GAPDH proteins were detected and performed ( D ) densitometry analysis Quantification; ( E ) siRNA 1 was pretransfected into A549 cells, and then the cells were infected with the virus at MOI = 0.5. Cell supernatants were collected at 4, 8, 12, 24 and 36 h after infection for TCID 50 assay to determine virus titer. ( F ) The SphK1-pRK5 (SphK1-Flag-pRK5) and Flag-pRK5 empty vector was pretransfected into A549 cells, then the cells were infected with the virus at MOI = 0.5, and the cells were collected for WB at 4, 8, 12, 24 and 36 h after infection. Detect the expression levels of SphK1, viral NP, M1 and GAPDH proteins and perform ( G ) densitometry analysis and quantification; ( H ) The constructed SphK1 eukaryotic expression vector was pretransfected into A549 cells, and then the cells were infected with the virus at MOI = 0.5. Cell supernatants were collected at 4, 8, 12, 24 and 36 h after infection for TCID 50 assay to determine virus titer. GAPDH was used as a loading control. Shown in the figure is the mean ± SD of the results of 3 trials. Differences were considered statistically significant at p < 0.05 (* p < 0.05, ** p < 0.01, ANOVA).

Article Snippet: siRNAs against SphK1 were transfected using Lipofectamine RNAiMax (Invitrogen, Carlsbad, CA, USA) at a final concentration of 20 nM following the manufacturer’s instructions. siRNAs against SphK1 (siRNA1: 5′-GCAGCUUCCUUGAACCAUUTT-3′, siRNA2: 5′-GAUUGCUGAUGUGGACCUTAA-3′, siRNA3: 5′-GUGCACCCAAACUACUUCUTT-3′) are constructed by GenePharma Corporation (Shanghai, China).

Techniques: Expressing, Negative Control, Transfection, Western Blot, Infection, Plasmid Preparation, Construct

Journal: Viruses

Article Title: Effect of the Interaction between Viral PB2 and Host SphK1 on H9N2 AIV Replication in Mammals

doi: 10.3390/v14071585

Figure Lengend Snippet: Interaction of SphK1 with PB2 proteins of different lineages. According to the plasmid transfection mode shown in the figure, 2 μg of the corresponding plasmids were transfected in the corresponding 293T cell wells. The cells were collected 24 h after transfection, immunoprecipitated with Flag antibody, and Western blotting was performed with HA and Flag antibody to detect whether ( A ) the DK1-like PB2 protein and ( B ) the F/98-like PB2 protein can interact with the SphK1 protein; ( C ) According to the plasmid transfection mode shown in the figure, 2 μg of the corresponding plasmids were transfected in the corresponding 293T cell wells. The cells were collected 24 h after transfection, and HA antibody was used for immunoprecipitation, and HA and Flag antibodies were used for Western blotting test to detect the interaction strength of SphK1 protein with F/98-like PB2 protein and DK1-like PB2 protein (IP: immunoprecipitation, IB: immunoblotting, Input: refer to whole cell lysates, Actin was used as a loading control).

Article Snippet: siRNAs against SphK1 were transfected using Lipofectamine RNAiMax (Invitrogen, Carlsbad, CA, USA) at a final concentration of 20 nM following the manufacturer’s instructions. siRNAs against SphK1 (siRNA1: 5′-GCAGCUUCCUUGAACCAUUTT-3′, siRNA2: 5′-GAUUGCUGAUGUGGACCUTAA-3′, siRNA3: 5′-GUGCACCCAAACUACUUCUTT-3′) are constructed by GenePharma Corporation (Shanghai, China).

Techniques: Plasmid Preparation, Transfection, Immunoprecipitation, Western Blot