Journal: Marine Drugs

Article Title: Recombinant N-acyl homoserine lactone-Lactonase AiiA QSI-1 Attenuates Aeromonas hydrophila Virulence Factors, Biofilm Formation and Reduces Mortality in Crucian Carp

doi: 10.3390/md17090499

Figure Lengend Snippet: Map of expression plasmid containing aiiA QSI-1 gene.

Article Snippet: The recombinant pET-30a- aiiA QSI-1 vector was then transformed into E. coli BL21(DE3) (Promega, Madison WI,, USA).

Techniques: Expressing, Plasmid Preparation

Journal: Marine Drugs

Article Title: Recombinant N-acyl homoserine lactone-Lactonase AiiA QSI-1 Attenuates Aeromonas hydrophila Virulence Factors, Biofilm Formation and Reduces Mortality in Crucian Carp

doi: 10.3390/md17090499

Figure Lengend Snippet: SDS-PAGE analysis of the recombinant AiiA QSI-1 . M: marker; 1: total protein of noninduced E. coli BL21(DE3)pET30a-aiiA QSI-1 cell lysate; 2: Supernatant of the sonication product of IPTG-induced E. coli BL21(DE3)pET30a-aiiA QSI-1 ; 3: Precipitate of the sonication product of IPTG-induced E. coli BL21(DE3)pET30a-aiiA QSI-1 ; 4: purified AiiA QSI-1 protein (as indicated by arrow).

Article Snippet: The recombinant pET-30a- aiiA QSI-1 vector was then transformed into E. coli BL21(DE3) (Promega, Madison WI,, USA).

Techniques: SDS Page, Recombinant, Marker, Sonication, Purification

Journal: Marine Drugs

Article Title: Recombinant N-acyl homoserine lactone-Lactonase AiiA QSI-1 Attenuates Aeromonas hydrophila Virulence Factors, Biofilm Formation and Reduces Mortality in Crucian Carp

doi: 10.3390/md17090499

Figure Lengend Snippet: AHL-degrading activity bioassays in vivo. Ten μL C6-HSL (10 μM) was mixed with 0 μL ( a ), 20 μL ( b ), 50 μL ( c ), 100 μL ( d ) AiiA QSI-1 , respectively and incubated at 30 °C.

Article Snippet: The recombinant pET-30a- aiiA QSI-1 vector was then transformed into E. coli BL21(DE3) (Promega, Madison WI,, USA).

Techniques: Activity Assay, In Vivo, Incubation

Journal: Marine Drugs

Article Title: Recombinant N-acyl homoserine lactone-Lactonase AiiA QSI-1 Attenuates Aeromonas hydrophila Virulence Factors, Biofilm Formation and Reduces Mortality in Crucian Carp

doi: 10.3390/md17090499

Figure Lengend Snippet: Effect of AiiA QSI-1 on the motility, production of virulence factors and biofilm, and growth of Aerononas hydrophila YJ-1. Swimming motility ( A ), Growth curve of planktonic bacteria ( B ), and biofilm production ( C ).

Article Snippet: The recombinant pET-30a- aiiA QSI-1 vector was then transformed into E. coli BL21(DE3) (Promega, Madison WI,, USA).

Techniques:

Journal: Marine Drugs

Article Title: Recombinant N-acyl homoserine lactone-Lactonase AiiA QSI-1 Attenuates Aeromonas hydrophila Virulence Factors, Biofilm Formation and Reduces Mortality in Crucian Carp

doi: 10.3390/md17090499

Figure Lengend Snippet: Effect of different concentrations of AiiA QSI-1 on A. hydrophila YJ-1 adherence and internalization to EPC. * p < 0.05; ** p < 0.01 in comparison to controls (no added QQ).

Article Snippet: The recombinant pET-30a- aiiA QSI-1 vector was then transformed into E. coli BL21(DE3) (Promega, Madison WI,, USA).

Techniques:

Journal: Marine Drugs

Article Title: Recombinant N-acyl homoserine lactone-Lactonase AiiA QSI-1 Attenuates Aeromonas hydrophila Virulence Factors, Biofilm Formation and Reduces Mortality in Crucian Carp

doi: 10.3390/md17090499

Figure Lengend Snippet: Percent survival of Crucian carp supplement with/without AiiA QSI-1 after Aeromonas hydrophila YJ-1 infection.

Article Snippet: The recombinant pET-30a- aiiA QSI-1 vector was then transformed into E. coli BL21(DE3) (Promega, Madison WI,, USA).

Techniques: Infection

Journal: Cancer Cell International

Article Title: LINC00115 promotes stemness and inhibits apoptosis of ovarian cancer stem cells by upregulating SOX9 and inhibiting the Wnt/β-catenin pathway through competitively binding to microRNA-30a

doi: 10.1186/s12935-021-02019-2

Figure Lengend Snippet: Sequences of 2 shRNAs of LINC00115

Article Snippet: The cultured CD133 + OCSCs were assigned into control shRNA (Ctrl), sh-LINC00115-1 (sh1), sh-LINC00115-2 (sh2), mimic-negative control (NC), miR-30a mimic, mimic-NC + vector-NC, miR-30a mimic + vector-NC, mimic-NC + SOX9-vector, miR-30a mimic + SOX9-vector, sh1 + inhibitor-NC, sh1 + miR-30a inhibitor, sh1 + vector-NC, sh1 + SOX9-vector, sh1 + PBS and sh1 + SKL2001 (a specific activator of the Wnt/β-catenin pathway) groups. miR-30a mimic, miR-30a inhibitor and their controls were purchased from Invitrogen Inc., SOX9-vector (the correct sequence of SOX9 was inserted into pCEP4 plasmid to construct pCEP4/SOX9 eukaryotic expression vector) and its NC (pCEP4 empty plasmid, named vector-NC), sh1, sh2 (two shRNA expression vectors, sh1 and sh2, were constructed using pRNAT-U6.1/neo plasmid) (Table ) and their controls (pRNAT-U6.1/neo empty plasmid, named Ctrl) were purchased from Guangzhou RiboBio Co., Ltd, (Guangzhou, Guangdong, China).

Techniques: Sequencing

Journal: Cancer Cell International

Article Title: LINC00115 promotes stemness and inhibits apoptosis of ovarian cancer stem cells by upregulating SOX9 and inhibiting the Wnt/β-catenin pathway through competitively binding to microRNA-30a

doi: 10.1186/s12935-021-02019-2

Figure Lengend Snippet: LINC00115 promotes the maintenance of OCSC stemness and inhibits its apoptosis. Two sh-LINC00115 vectors (sh1 and sh2) and their negative control (Ctrl) were used to transfect CD133 + OCSCs cells. A The expression heat map of 50 DEGs/lncRNAs on GSE66957, in which Y-axis represents DEGs/lncRNAs. The upper right histogram is the color order, and each rectangle corresponds to a sample expression value; B Relative expression of LINC00115 in A2780 cells and CD133 + OCSCs detected by RT-qPCR, compared with the A2780 cells, * p < 0.05; C The sphere-forming rate of CD133 + OCSCs was observed under a microscope, VS the Ctrl group, *** p < 0.001; D , E . Relative expression of LINC00115, CD133, CD44, NANOG and POU5F1 in CD133 + OCSCs was detected using RT-qPCR and Western blot analysis, vs the Ctrl group, * p < 0.05; F Cell apoptosis was measured using flow cytometry; G. Relative Caspase-3 activity in cells of each group, VS the Ctrl group, * p < 0.05; H. Percentage of DNA damage in cell supernatant of each group, VS the Ctrl group, * p < 0.05. Data in panel B were analyzed using the unpaired t test, and the data in panels C-H were analyzed using one-way ANOVA and Tukey’s multiple comparison test. The experiment was performed in triplicate and data were expressed as means ± standard deviation

Article Snippet: The cultured CD133 + OCSCs were assigned into control shRNA (Ctrl), sh-LINC00115-1 (sh1), sh-LINC00115-2 (sh2), mimic-negative control (NC), miR-30a mimic, mimic-NC + vector-NC, miR-30a mimic + vector-NC, mimic-NC + SOX9-vector, miR-30a mimic + SOX9-vector, sh1 + inhibitor-NC, sh1 + miR-30a inhibitor, sh1 + vector-NC, sh1 + SOX9-vector, sh1 + PBS and sh1 + SKL2001 (a specific activator of the Wnt/β-catenin pathway) groups. miR-30a mimic, miR-30a inhibitor and their controls were purchased from Invitrogen Inc., SOX9-vector (the correct sequence of SOX9 was inserted into pCEP4 plasmid to construct pCEP4/SOX9 eukaryotic expression vector) and its NC (pCEP4 empty plasmid, named vector-NC), sh1, sh2 (two shRNA expression vectors, sh1 and sh2, were constructed using pRNAT-U6.1/neo plasmid) (Table ) and their controls (pRNAT-U6.1/neo empty plasmid, named Ctrl) were purchased from Guangzhou RiboBio Co., Ltd, (Guangzhou, Guangdong, China).

Techniques: Negative Control, Expressing, Quantitative RT-PCR, Microscopy, Western Blot, Flow Cytometry, Activity Assay, Comparison, Standard Deviation

Journal: Cancer Cell International

Article Title: LINC00115 promotes stemness and inhibits apoptosis of ovarian cancer stem cells by upregulating SOX9 and inhibiting the Wnt/β-catenin pathway through competitively binding to microRNA-30a

doi: 10.1186/s12935-021-02019-2

Figure Lengend Snippet: LINC00115 participates in the maintenance and apoptosis of OCSCs through the miR-30a/SOX9 axis. A Microscopic observation of spheres (100 ×) and sphere-forming rate of OCSCs measured by sphere-forming assay; B , C Relative levels of CD133, SOX9, CD44, NANOG, and POU5F1 were detected by RT-qPCR and Western blot analysis; D Cell apoptosis was measured by flow cytometry; E Relative Caspase-3 activity; F Relative percentage of DNA damage in cell supernatant of each group. VS the sh1 + inhibitor-NC group, * p < 0.05; vs the sh1 + vector-NC group, # p < 0.05. Data were analyzed using one-way ANOVA and Tukey’s multiple comparison test. The experiment was performed in triplicate and data were expressed as means ± standard deviation

Article Snippet: The cultured CD133 + OCSCs were assigned into control shRNA (Ctrl), sh-LINC00115-1 (sh1), sh-LINC00115-2 (sh2), mimic-negative control (NC), miR-30a mimic, mimic-NC + vector-NC, miR-30a mimic + vector-NC, mimic-NC + SOX9-vector, miR-30a mimic + SOX9-vector, sh1 + inhibitor-NC, sh1 + miR-30a inhibitor, sh1 + vector-NC, sh1 + SOX9-vector, sh1 + PBS and sh1 + SKL2001 (a specific activator of the Wnt/β-catenin pathway) groups. miR-30a mimic, miR-30a inhibitor and their controls were purchased from Invitrogen Inc., SOX9-vector (the correct sequence of SOX9 was inserted into pCEP4 plasmid to construct pCEP4/SOX9 eukaryotic expression vector) and its NC (pCEP4 empty plasmid, named vector-NC), sh1, sh2 (two shRNA expression vectors, sh1 and sh2, were constructed using pRNAT-U6.1/neo plasmid) (Table ) and their controls (pRNAT-U6.1/neo empty plasmid, named Ctrl) were purchased from Guangzhou RiboBio Co., Ltd, (Guangzhou, Guangdong, China).

Techniques: Quantitative RT-PCR, Western Blot, Flow Cytometry, Activity Assay, Plasmid Preparation, Comparison, Standard Deviation

Journal: Cancer Cell International

Article Title: LINC00115 promotes stemness and inhibits apoptosis of ovarian cancer stem cells by upregulating SOX9 and inhibiting the Wnt/β-catenin pathway through competitively binding to microRNA-30a

doi: 10.1186/s12935-021-02019-2

Figure Lengend Snippet: Wnt/β-catenin pathway participates in the maintenance of OCSCs. OCSCs stably transfected with sh1 were delivered miR-30a inhibitor or SOX9-vector, or 5 mM Wnt/β-catenin specific activator SKL2001. A , B . Western blot analysis detected the expression of Wnt/β-catenin pathway; C. Microscopic observation of spheres (100 ×) and sphere-forming rate of OCSCs measured by sphere-forming assay; D Relative levels of CD133, SOX9, CD44, NANOG, and POU5F1 in CD133 + OCSCs were measured using Western blot analysis; E Cell apoptosis was measured by flow cytometry; F Relative Caspase-3 activity; G. Relative percentage of DNA damage in cell supernatant of each group. VS the sh1 + PBS group, * p < 0.05. Data in panels A and B were analyzed using one-way ANOVA and Tukey’s multiple comparison test, while data in panels C and F were analyzed using the unpaired t test. The experiment was performed in triplicate and data were expressed as means ± standard deviation

Article Snippet: The cultured CD133 + OCSCs were assigned into control shRNA (Ctrl), sh-LINC00115-1 (sh1), sh-LINC00115-2 (sh2), mimic-negative control (NC), miR-30a mimic, mimic-NC + vector-NC, miR-30a mimic + vector-NC, mimic-NC + SOX9-vector, miR-30a mimic + SOX9-vector, sh1 + inhibitor-NC, sh1 + miR-30a inhibitor, sh1 + vector-NC, sh1 + SOX9-vector, sh1 + PBS and sh1 + SKL2001 (a specific activator of the Wnt/β-catenin pathway) groups. miR-30a mimic, miR-30a inhibitor and their controls were purchased from Invitrogen Inc., SOX9-vector (the correct sequence of SOX9 was inserted into pCEP4 plasmid to construct pCEP4/SOX9 eukaryotic expression vector) and its NC (pCEP4 empty plasmid, named vector-NC), sh1, sh2 (two shRNA expression vectors, sh1 and sh2, were constructed using pRNAT-U6.1/neo plasmid) (Table ) and their controls (pRNAT-U6.1/neo empty plasmid, named Ctrl) were purchased from Guangzhou RiboBio Co., Ltd, (Guangzhou, Guangdong, China).

Techniques: Stable Transfection, Transfection, Plasmid Preparation, Western Blot, Expressing, Flow Cytometry, Activity Assay, Comparison, Standard Deviation

Journal: Cancer Cell International

Article Title: LINC00115 promotes stemness and inhibits apoptosis of ovarian cancer stem cells by upregulating SOX9 and inhibiting the Wnt/β-catenin pathway through competitively binding to microRNA-30a

doi: 10.1186/s12935-021-02019-2

Figure Lengend Snippet: LINC00115 promotes OCSC growth in vivo. Nude mice were randomly assigned into three groups, 6 mice in each group, namely Ctrl group (injected with OCSCs), sh1 group (injected with sh1-treated OCSCs) and sh2 group (injected with sh2-treated OCSCs). A – C In vivo OCSC growth (including tumor size and weight) in nude mice transplanted with tumor cells; D Immunohistochemical staining of Ki67. Data were analyzed using one-way ANOVA and Tukey’s multiple comparison test. VS the Ctrl group, * p < 0.05; ** p < 0.01

Article Snippet: The cultured CD133 + OCSCs were assigned into control shRNA (Ctrl), sh-LINC00115-1 (sh1), sh-LINC00115-2 (sh2), mimic-negative control (NC), miR-30a mimic, mimic-NC + vector-NC, miR-30a mimic + vector-NC, mimic-NC + SOX9-vector, miR-30a mimic + SOX9-vector, sh1 + inhibitor-NC, sh1 + miR-30a inhibitor, sh1 + vector-NC, sh1 + SOX9-vector, sh1 + PBS and sh1 + SKL2001 (a specific activator of the Wnt/β-catenin pathway) groups. miR-30a mimic, miR-30a inhibitor and their controls were purchased from Invitrogen Inc., SOX9-vector (the correct sequence of SOX9 was inserted into pCEP4 plasmid to construct pCEP4/SOX9 eukaryotic expression vector) and its NC (pCEP4 empty plasmid, named vector-NC), sh1, sh2 (two shRNA expression vectors, sh1 and sh2, were constructed using pRNAT-U6.1/neo plasmid) (Table ) and their controls (pRNAT-U6.1/neo empty plasmid, named Ctrl) were purchased from Guangzhou RiboBio Co., Ltd, (Guangzhou, Guangdong, China).

Techniques: In Vivo, Injection, Immunohistochemical staining, Staining, Comparison

Journal: Cancer Cell International

Article Title: LINC00115 promotes stemness and inhibits apoptosis of ovarian cancer stem cells by upregulating SOX9 and inhibiting the Wnt/β-catenin pathway through competitively binding to microRNA-30a

doi: 10.1186/s12935-021-02019-2

Figure Lengend Snippet: Sequences of 2 shRNAs of LINC00115

Article Snippet: The cultured CD133 + OCSCs were assigned into control shRNA (Ctrl), sh-LINC00115-1 (sh1), sh-LINC00115-2 (sh2), mimic-negative control (NC), miR-30a mimic, mimic-NC + vector-NC, miR-30a mimic + vector-NC, mimic-NC + SOX9-vector, miR-30a mimic + SOX9-vector, sh1 + inhibitor-NC, sh1 + miR-30a inhibitor, sh1 + vector-NC, sh1 + SOX9-vector, sh1 + PBS and sh1 + SKL2001 (a specific activator of the Wnt/β-catenin pathway) groups. miR-30a mimic, miR-30a inhibitor and their controls were purchased from Invitrogen Inc., SOX9-vector (the correct sequence of SOX9 was inserted into pCEP4 plasmid to construct pCEP4/SOX9 eukaryotic expression vector) and its NC (pCEP4 empty plasmid, named vector-NC), sh1, sh2 (two shRNA expression vectors, sh1 and sh2, were constructed using pRNAT-U6.1/neo plasmid) (Table ) and their controls (pRNAT-U6.1/neo empty plasmid, named Ctrl) were purchased from Guangzhou RiboBio Co., Ltd, (Guangzhou, Guangdong, China).

Techniques: Sequencing

Journal: Cancer Cell International

Article Title: LINC00115 promotes stemness and inhibits apoptosis of ovarian cancer stem cells by upregulating SOX9 and inhibiting the Wnt/β-catenin pathway through competitively binding to microRNA-30a

doi: 10.1186/s12935-021-02019-2

Figure Lengend Snippet: LINC00115 promotes the maintenance of OCSC stemness and inhibits its apoptosis. Two sh-LINC00115 vectors (sh1 and sh2) and their negative control (Ctrl) were used to transfect CD133 + OCSCs cells. A The expression heat map of 50 DEGs/lncRNAs on GSE66957, in which Y-axis represents DEGs/lncRNAs. The upper right histogram is the color order, and each rectangle corresponds to a sample expression value; B Relative expression of LINC00115 in A2780 cells and CD133 + OCSCs detected by RT-qPCR, compared with the A2780 cells, * p < 0.05; C The sphere-forming rate of CD133 + OCSCs was observed under a microscope, VS the Ctrl group, *** p < 0.001; D , E . Relative expression of LINC00115, CD133, CD44, NANOG and POU5F1 in CD133 + OCSCs was detected using RT-qPCR and Western blot analysis, vs the Ctrl group, * p < 0.05; F Cell apoptosis was measured using flow cytometry; G. Relative Caspase-3 activity in cells of each group, VS the Ctrl group, * p < 0.05; H. Percentage of DNA damage in cell supernatant of each group, VS the Ctrl group, * p < 0.05. Data in panel B were analyzed using the unpaired t test, and the data in panels C-H were analyzed using one-way ANOVA and Tukey’s multiple comparison test. The experiment was performed in triplicate and data were expressed as means ± standard deviation

Article Snippet: The cultured CD133 + OCSCs were assigned into control shRNA (Ctrl), sh-LINC00115-1 (sh1), sh-LINC00115-2 (sh2), mimic-negative control (NC), miR-30a mimic, mimic-NC + vector-NC, miR-30a mimic + vector-NC, mimic-NC + SOX9-vector, miR-30a mimic + SOX9-vector, sh1 + inhibitor-NC, sh1 + miR-30a inhibitor, sh1 + vector-NC, sh1 + SOX9-vector, sh1 + PBS and sh1 + SKL2001 (a specific activator of the Wnt/β-catenin pathway) groups. miR-30a mimic, miR-30a inhibitor and their controls were purchased from Invitrogen Inc., SOX9-vector (the correct sequence of SOX9 was inserted into pCEP4 plasmid to construct pCEP4/SOX9 eukaryotic expression vector) and its NC (pCEP4 empty plasmid, named vector-NC), sh1, sh2 (two shRNA expression vectors, sh1 and sh2, were constructed using pRNAT-U6.1/neo plasmid) (Table ) and their controls (pRNAT-U6.1/neo empty plasmid, named Ctrl) were purchased from Guangzhou RiboBio Co., Ltd, (Guangzhou, Guangdong, China).

Techniques: Negative Control, Expressing, Quantitative RT-PCR, Microscopy, Western Blot, Flow Cytometry, Activity Assay, Comparison, Standard Deviation

Journal: Cancer Cell International

Article Title: LINC00115 promotes stemness and inhibits apoptosis of ovarian cancer stem cells by upregulating SOX9 and inhibiting the Wnt/β-catenin pathway through competitively binding to microRNA-30a

doi: 10.1186/s12935-021-02019-2

Figure Lengend Snippet: LINC00115 participates in the maintenance and apoptosis of OCSCs through the miR-30a/SOX9 axis. A Microscopic observation of spheres (100 ×) and sphere-forming rate of OCSCs measured by sphere-forming assay; B , C Relative levels of CD133, SOX9, CD44, NANOG, and POU5F1 were detected by RT-qPCR and Western blot analysis; D Cell apoptosis was measured by flow cytometry; E Relative Caspase-3 activity; F Relative percentage of DNA damage in cell supernatant of each group. VS the sh1 + inhibitor-NC group, * p < 0.05; vs the sh1 + vector-NC group, # p < 0.05. Data were analyzed using one-way ANOVA and Tukey’s multiple comparison test. The experiment was performed in triplicate and data were expressed as means ± standard deviation

Article Snippet: The cultured CD133 + OCSCs were assigned into control shRNA (Ctrl), sh-LINC00115-1 (sh1), sh-LINC00115-2 (sh2), mimic-negative control (NC), miR-30a mimic, mimic-NC + vector-NC, miR-30a mimic + vector-NC, mimic-NC + SOX9-vector, miR-30a mimic + SOX9-vector, sh1 + inhibitor-NC, sh1 + miR-30a inhibitor, sh1 + vector-NC, sh1 + SOX9-vector, sh1 + PBS and sh1 + SKL2001 (a specific activator of the Wnt/β-catenin pathway) groups. miR-30a mimic, miR-30a inhibitor and their controls were purchased from Invitrogen Inc., SOX9-vector (the correct sequence of SOX9 was inserted into pCEP4 plasmid to construct pCEP4/SOX9 eukaryotic expression vector) and its NC (pCEP4 empty plasmid, named vector-NC), sh1, sh2 (two shRNA expression vectors, sh1 and sh2, were constructed using pRNAT-U6.1/neo plasmid) (Table ) and their controls (pRNAT-U6.1/neo empty plasmid, named Ctrl) were purchased from Guangzhou RiboBio Co., Ltd, (Guangzhou, Guangdong, China).

Techniques: Quantitative RT-PCR, Western Blot, Flow Cytometry, Activity Assay, Plasmid Preparation, Comparison, Standard Deviation

Journal: Cancer Cell International

Article Title: LINC00115 promotes stemness and inhibits apoptosis of ovarian cancer stem cells by upregulating SOX9 and inhibiting the Wnt/β-catenin pathway through competitively binding to microRNA-30a

doi: 10.1186/s12935-021-02019-2

Figure Lengend Snippet: Wnt/β-catenin pathway participates in the maintenance of OCSCs. OCSCs stably transfected with sh1 were delivered miR-30a inhibitor or SOX9-vector, or 5 mM Wnt/β-catenin specific activator SKL2001. A , B . Western blot analysis detected the expression of Wnt/β-catenin pathway; C. Microscopic observation of spheres (100 ×) and sphere-forming rate of OCSCs measured by sphere-forming assay; D Relative levels of CD133, SOX9, CD44, NANOG, and POU5F1 in CD133 + OCSCs were measured using Western blot analysis; E Cell apoptosis was measured by flow cytometry; F Relative Caspase-3 activity; G. Relative percentage of DNA damage in cell supernatant of each group. VS the sh1 + PBS group, * p < 0.05. Data in panels A and B were analyzed using one-way ANOVA and Tukey’s multiple comparison test, while data in panels C and F were analyzed using the unpaired t test. The experiment was performed in triplicate and data were expressed as means ± standard deviation

Article Snippet: The cultured CD133 + OCSCs were assigned into control shRNA (Ctrl), sh-LINC00115-1 (sh1), sh-LINC00115-2 (sh2), mimic-negative control (NC), miR-30a mimic, mimic-NC + vector-NC, miR-30a mimic + vector-NC, mimic-NC + SOX9-vector, miR-30a mimic + SOX9-vector, sh1 + inhibitor-NC, sh1 + miR-30a inhibitor, sh1 + vector-NC, sh1 + SOX9-vector, sh1 + PBS and sh1 + SKL2001 (a specific activator of the Wnt/β-catenin pathway) groups. miR-30a mimic, miR-30a inhibitor and their controls were purchased from Invitrogen Inc., SOX9-vector (the correct sequence of SOX9 was inserted into pCEP4 plasmid to construct pCEP4/SOX9 eukaryotic expression vector) and its NC (pCEP4 empty plasmid, named vector-NC), sh1, sh2 (two shRNA expression vectors, sh1 and sh2, were constructed using pRNAT-U6.1/neo plasmid) (Table ) and their controls (pRNAT-U6.1/neo empty plasmid, named Ctrl) were purchased from Guangzhou RiboBio Co., Ltd, (Guangzhou, Guangdong, China).

Techniques: Stable Transfection, Transfection, Plasmid Preparation, Western Blot, Expressing, Flow Cytometry, Activity Assay, Comparison, Standard Deviation

Journal: Cancer Cell International

Article Title: LINC00115 promotes stemness and inhibits apoptosis of ovarian cancer stem cells by upregulating SOX9 and inhibiting the Wnt/β-catenin pathway through competitively binding to microRNA-30a

doi: 10.1186/s12935-021-02019-2

Figure Lengend Snippet: LINC00115 promotes OCSC growth in vivo. Nude mice were randomly assigned into three groups, 6 mice in each group, namely Ctrl group (injected with OCSCs), sh1 group (injected with sh1-treated OCSCs) and sh2 group (injected with sh2-treated OCSCs). A – C In vivo OCSC growth (including tumor size and weight) in nude mice transplanted with tumor cells; D Immunohistochemical staining of Ki67. Data were analyzed using one-way ANOVA and Tukey’s multiple comparison test. VS the Ctrl group, * p < 0.05; ** p < 0.01

Article Snippet: The cultured CD133 + OCSCs were assigned into control shRNA (Ctrl), sh-LINC00115-1 (sh1), sh-LINC00115-2 (sh2), mimic-negative control (NC), miR-30a mimic, mimic-NC + vector-NC, miR-30a mimic + vector-NC, mimic-NC + SOX9-vector, miR-30a mimic + SOX9-vector, sh1 + inhibitor-NC, sh1 + miR-30a inhibitor, sh1 + vector-NC, sh1 + SOX9-vector, sh1 + PBS and sh1 + SKL2001 (a specific activator of the Wnt/β-catenin pathway) groups. miR-30a mimic, miR-30a inhibitor and their controls were purchased from Invitrogen Inc., SOX9-vector (the correct sequence of SOX9 was inserted into pCEP4 plasmid to construct pCEP4/SOX9 eukaryotic expression vector) and its NC (pCEP4 empty plasmid, named vector-NC), sh1, sh2 (two shRNA expression vectors, sh1 and sh2, were constructed using pRNAT-U6.1/neo plasmid) (Table ) and their controls (pRNAT-U6.1/neo empty plasmid, named Ctrl) were purchased from Guangzhou RiboBio Co., Ltd, (Guangzhou, Guangdong, China).

Techniques: In Vivo, Injection, Immunohistochemical staining, Staining, Comparison