Journal: Cancer Cell International

Article Title: Enhanced osteopontin splicing regulated by RUNX2 is HDAC-dependent and induces invasive phenotypes in NSCLC cells

doi: 10.1186/s12935-019-1033-5

Figure Lengend Snippet: Primers for PCR-based assays and siRNAs used for transfection

Article Snippet: The expression cassettes of OPN-SIs (OPNa, OPNb and OPNc) and RUNX2 were cloned individually into pENTER vectors by Vigene Bioscience Co., Ltd. (Shandong, China) and amplified in bacterial host cells.

Techniques: Sequencing

Journal: Cancer Cell International

Article Title: Enhanced osteopontin splicing regulated by RUNX2 is HDAC-dependent and induces invasive phenotypes in NSCLC cells

doi: 10.1186/s12935-019-1033-5

Figure Lengend Snippet: Upregulation of RUNX2 associated with increased OPN expression during the EMT progression in TGF-β treated A549 cells. a TGF-β treatments for 48 h increased cell migration. b TGF-β treatments induced significant changes in the expression of major EMT marker genes at both mRNA (left) and protein (right) levels. c The mRNA (upper) and protein (lower) expression of OPN and RUNX2 were synchronically increased following TGF-β treatments

Article Snippet: The expression cassettes of OPN-SIs (OPNa, OPNb and OPNc) and RUNX2 were cloned individually into pENTER vectors by Vigene Bioscience Co., Ltd. (Shandong, China) and amplified in bacterial host cells.

Techniques: Expressing, Migration, Marker

Journal: Cancer Cell International

Article Title: Enhanced osteopontin splicing regulated by RUNX2 is HDAC-dependent and induces invasive phenotypes in NSCLC cells

doi: 10.1186/s12935-019-1033-5

Figure Lengend Snippet: RUNX2 overexpression enhanced the expression of OPN, especially of splicing isoform c. a Schematic illustration of isoform-specific primers used for the quantitative analyses of OPN-SIs. b TGF-β induction increased the opn transcript levels unproportionally among different splicing isoforms. c TGF-β preferentially promoted OPNc splicing in A549 cells. d Western blot of RUNX2 in A549 cells transfected with an overexpression plasmid. e Overexpression of RUNX2 altered the OPN-SIs and OPNt mRNA levels. f RUNX2 overexpression selectively increased OPNc transcript levels. g Knockdown of RUNX2 expression attenuated TGF-β induced opn expression in A549 cells

Article Snippet: The expression cassettes of OPN-SIs (OPNa, OPNb and OPNc) and RUNX2 were cloned individually into pENTER vectors by Vigene Bioscience Co., Ltd. (Shandong, China) and amplified in bacterial host cells.

Techniques: Over Expression, Expressing, Western Blot, Transfection, Plasmid Preparation, Knockdown

Journal: Cancer Cell International

Article Title: Enhanced osteopontin splicing regulated by RUNX2 is HDAC-dependent and induces invasive phenotypes in NSCLC cells

doi: 10.1186/s12935-019-1033-5

Figure Lengend Snippet: RUNX2 dependent OPNc splicing required normal activities of HDAC1 or HDAC2. a Treatment of A549 cells with HDACs inhibitor TSA suppressed OPNc splicing induced by RUNX2 overexpression. b Western blot of RUNX2 and OPN in RUNX2 overexpressed A549 cells following TSA treatment. c Inhibition of HDAC1 activity by NaB deprived RUNX2 induced OPNc splicing. d Western blot of RUNX2 and OPN in RUNX2 overexpressed A549 cells treated with NaB. e Knockdown of HDAC1 or HDAC2, but not HDAC3, decreased RUNX2-induced OPNc splicing. f Western blots of HDAC1, HDAC2 and HDAC3 from A549 cells transfected with the targeted siRNAs

Article Snippet: The expression cassettes of OPN-SIs (OPNa, OPNb and OPNc) and RUNX2 were cloned individually into pENTER vectors by Vigene Bioscience Co., Ltd. (Shandong, China) and amplified in bacterial host cells.

Techniques: Over Expression, Western Blot, Inhibition, Activity Assay, Knockdown, Transfection

Journal: Cancer Cell International

Article Title: Enhanced osteopontin splicing regulated by RUNX2 is HDAC-dependent and induces invasive phenotypes in NSCLC cells

doi: 10.1186/s12935-019-1033-5

Figure Lengend Snippet: TGF-β induced OPNc expression enhanced the mobility of SK-MES-1 cells with a dependence to HDACs. a TGF-β treatments for 48 h increased the invasiveness of SK-MES-1 cells. b TGF-β induction increased OPNt and OPN-SIs levels with a preference to OPNc. c TGF-β promoted OPNc splicing with most significance in SK-MES-1 cells. d Knockdown of HDAC1 or HDAC2 significantly decreased the RUNX2-induced OPNc splicing. e The HDAC1 and HDAC2 expression were markedly reduced in the SK-MES-1 cells transfected with the targeted siRNAs. f Overexpression of OPN-SIs significantly promoted the migration of SK-MES-1 cells

Article Snippet: The expression cassettes of OPN-SIs (OPNa, OPNb and OPNc) and RUNX2 were cloned individually into pENTER vectors by Vigene Bioscience Co., Ltd. (Shandong, China) and amplified in bacterial host cells.

Techniques: Expressing, Knockdown, Transfection, Over Expression, Migration

Journal: Cancer Cell International

Article Title: Enhanced osteopontin splicing regulated by RUNX2 is HDAC-dependent and induces invasive phenotypes in NSCLC cells

doi: 10.1186/s12935-019-1033-5

Figure Lengend Snippet: Primers for PCR-based assays and siRNAs used for transfection

Article Snippet: A set of minigene reporters for the splicing of OPNb and OPNc was constructed by Genechem Co., Ltd. (Shanghai, China).

Techniques: Sequencing

Journal: Cancer Cell International

Article Title: Enhanced osteopontin splicing regulated by RUNX2 is HDAC-dependent and induces invasive phenotypes in NSCLC cells

doi: 10.1186/s12935-019-1033-5

Figure Lengend Snippet: RUNX2 overexpression enhanced the expression of OPN, especially of splicing isoform c. a Schematic illustration of isoform-specific primers used for the quantitative analyses of OPN-SIs. b TGF-β induction increased the opn transcript levels unproportionally among different splicing isoforms. c TGF-β preferentially promoted OPNc splicing in A549 cells. d Western blot of RUNX2 in A549 cells transfected with an overexpression plasmid. e Overexpression of RUNX2 altered the OPN-SIs and OPNt mRNA levels. f RUNX2 overexpression selectively increased OPNc transcript levels. g Knockdown of RUNX2 expression attenuated TGF-β induced opn expression in A549 cells

Article Snippet: A set of minigene reporters for the splicing of OPNb and OPNc was constructed by Genechem Co., Ltd. (Shanghai, China).

Techniques: Over Expression, Expressing, Western Blot, Transfection, Plasmid Preparation, Knockdown

Journal: Cancer Cell International

Article Title: Enhanced osteopontin splicing regulated by RUNX2 is HDAC-dependent and induces invasive phenotypes in NSCLC cells

doi: 10.1186/s12935-019-1033-5

Figure Lengend Snippet: Knock down of SRSF1 reduced mRNA levels of OPN-SIs. a RNAi of SRSF1 decreased OPNc levels in A549 cells. b Inhibition of OPNb and OPNc splicing from reporter assays following SRSF1 siRNA transfection. c Western blot of SRSF1 in A549 cells transfected with targeted siRNAs

Article Snippet: A set of minigene reporters for the splicing of OPNb and OPNc was constructed by Genechem Co., Ltd. (Shanghai, China).

Techniques: Knockdown, Inhibition, Transfection, Western Blot

Journal: Cancer Cell International

Article Title: Enhanced osteopontin splicing regulated by RUNX2 is HDAC-dependent and induces invasive phenotypes in NSCLC cells

doi: 10.1186/s12935-019-1033-5

Figure Lengend Snippet: RUNX2 dependent OPNc splicing required normal activities of HDAC1 or HDAC2. a Treatment of A549 cells with HDACs inhibitor TSA suppressed OPNc splicing induced by RUNX2 overexpression. b Western blot of RUNX2 and OPN in RUNX2 overexpressed A549 cells following TSA treatment. c Inhibition of HDAC1 activity by NaB deprived RUNX2 induced OPNc splicing. d Western blot of RUNX2 and OPN in RUNX2 overexpressed A549 cells treated with NaB. e Knockdown of HDAC1 or HDAC2, but not HDAC3, decreased RUNX2-induced OPNc splicing. f Western blots of HDAC1, HDAC2 and HDAC3 from A549 cells transfected with the targeted siRNAs

Article Snippet: A set of minigene reporters for the splicing of OPNb and OPNc was constructed by Genechem Co., Ltd. (Shanghai, China).

Techniques: Over Expression, Western Blot, Inhibition, Activity Assay, Knockdown, Transfection

Journal: Cancer Cell International

Article Title: Enhanced osteopontin splicing regulated by RUNX2 is HDAC-dependent and induces invasive phenotypes in NSCLC cells

doi: 10.1186/s12935-019-1033-5

Figure Lengend Snippet: OPNc overexpression was most potent to induce an invasive phenotype in A549 cells among OPN-SIs. a Overexpression of OPNb and OPNc significantly increased the invasiveness of transfected cells. b Overexpression of OPN-SIs promoted cell mobility and migration. c Immunoblots of EMT marker proteins in cells with overexpression of OPN-SIs

Article Snippet: A set of minigene reporters for the splicing of OPNb and OPNc was constructed by Genechem Co., Ltd. (Shanghai, China).

Techniques: Over Expression, Transfection, Migration, Western Blot, Marker

Journal: Cancer Cell International

Article Title: Enhanced osteopontin splicing regulated by RUNX2 is HDAC-dependent and induces invasive phenotypes in NSCLC cells

doi: 10.1186/s12935-019-1033-5

Figure Lengend Snippet: TGF-β induced OPNc expression enhanced the mobility of SK-MES-1 cells with a dependence to HDACs. a TGF-β treatments for 48 h increased the invasiveness of SK-MES-1 cells. b TGF-β induction increased OPNt and OPN-SIs levels with a preference to OPNc. c TGF-β promoted OPNc splicing with most significance in SK-MES-1 cells. d Knockdown of HDAC1 or HDAC2 significantly decreased the RUNX2-induced OPNc splicing. e The HDAC1 and HDAC2 expression were markedly reduced in the SK-MES-1 cells transfected with the targeted siRNAs. f Overexpression of OPN-SIs significantly promoted the migration of SK-MES-1 cells

Article Snippet: A set of minigene reporters for the splicing of OPNb and OPNc was constructed by Genechem Co., Ltd. (Shanghai, China).

Techniques: Expressing, Knockdown, Transfection, Over Expression, Migration