Journal: International Journal of Oral Science

Article Title: Identification of the involvement of LOXL4 in generation of keratocystic odontogenic tumors by RNA-Seq analysis

doi: 10.1038/ijos.2013.96

Figure Lengend Snippet: Genes identified as up-regulated or down-regulated by RNA-Seq and microarray analysis

Article Snippet: Human LOXL4 cDNA was purchased from Sino Biological and cloned in pCMV/hygro (CV004; Sino Biological, Beijng, China) for transient overexpression with a DNA Ligation Kit (2011A; TAKARA, Kyoto, Japan).

Techniques: Microarray

Journal: International Journal of Oral Science

Article Title: Identification of the involvement of LOXL4 in generation of keratocystic odontogenic tumors by RNA-Seq analysis

doi: 10.1038/ijos.2013.96

Figure Lengend Snippet: Candidate genes possibly involved in keratocystic odontogenic tumor progression

Article Snippet: Human LOXL4 cDNA was purchased from Sino Biological and cloned in pCMV/hygro (CV004; Sino Biological, Beijng, China) for transient overexpression with a DNA Ligation Kit (2011A; TAKARA, Kyoto, Japan).

Techniques: Microarray

Journal: International Journal of Oral Science

Article Title: Identification of the involvement of LOXL4 in generation of keratocystic odontogenic tumors by RNA-Seq analysis

doi: 10.1038/ijos.2013.96

Figure Lengend Snippet: Confirmation of RNA-Seq results. ( a ) QPCR analysis of TCTA, LARP6, RGS12 and LOXL4 mRNA expression in primary stromal fibroblasts of KCOT, DC and healthy GG. The data were consistent with the RNA-Seq and microarray results. ( b ) QPCR analysis of LOXL4 mRNA expression in primary stromal fibroblasts of KCOT, DC and GG. Each bar corresponds to the mean±s.d. of three independent experiments performed in triplicate with three technical repeats. * P <0.05, ** P <0.001. DC, dentigerous cysts; GG, gingival primary stromal fibroblast cells; KCOT, keratocystic odontogenic tumors; LARP6, La ribonucleoprotein domain family, member 6; LOXL4, lysyl oxidase-like 4; QPCR, quantitative PCR; RGS12, regulator of G-protein signaling 12; RNA-Seq, RNA sequencing; TCTA, T-cell leukemia translocation-associated gene.

Article Snippet: Human LOXL4 cDNA was purchased from Sino Biological and cloned in pCMV/hygro (CV004; Sino Biological, Beijng, China) for transient overexpression with a DNA Ligation Kit (2011A; TAKARA, Kyoto, Japan).

Techniques: RNA Sequencing Assay, Expressing, Microarray, Real-time Polymerase Chain Reaction, Translocation Assay

Journal: International Journal of Oral Science

Article Title: Identification of the involvement of LOXL4 in generation of keratocystic odontogenic tumors by RNA-Seq analysis

doi: 10.1038/ijos.2013.96

Figure Lengend Snippet: LOXL4 is overexpressed in the stroma of KCOT. ( a ) LOXL4 concentration in the supernatants of primary fibroblasts derived from KCOT, GG and DC ( n =3 per tissue) by ELISA. ( b ) LOXL4 concentration in the cell lysates of primary fibroblasts derived from KCOT, GG and DC ( n =3 per tissue) by ELISA. Data are the means of triplicates from a representative experiment±s.d.; * P <0.05, ** P <0.001. DC, dentigerous cysts; ELISA, enzyme-linked immunosorbent assay; GG, gingival primary stromal fibroblast cells; KCOT, keratocystic odontogenic tumors; LOXL4, lysyl oxidase-like 4.

Article Snippet: Human LOXL4 cDNA was purchased from Sino Biological and cloned in pCMV/hygro (CV004; Sino Biological, Beijng, China) for transient overexpression with a DNA Ligation Kit (2011A; TAKARA, Kyoto, Japan).

Techniques: Concentration Assay, Derivative Assay, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Oral Science

Article Title: Identification of the involvement of LOXL4 in generation of keratocystic odontogenic tumors by RNA-Seq analysis

doi: 10.1038/ijos.2013.96

Figure Lengend Snippet: Effects of LOXL4 on angiogenesis. ( a ) LOXL4 expression and microvessel density in DC, GG and KCOT stroma. Immunohistochemical analysis of LOXL4 ( a1 , a2 , a4 , a5 ). Few LOXL4-positive fibroblasts were observed in GG ( a1 , a2 ) or DC ( a4 , a5 ). ( a7 , a8 ) Most of the KCOT stromal fibroblasts were strongly LOXL4-positive. ( a3 , a6 ) Few CD105-positive microvessels were observed adjacent to the epithelium of normal GG ( a3 ) and dentigerous cysts ( a6 ). A high density of strongly CD105-positive microvessels was found in the stroma immediately beneath the epithelium lining KCOT ( a9 ). The intensity of LOXL4 protein expression was consistent with the density of CD105-positive microvessels in GG ( a3 ), DC ( a6 ) and KCOT ( a9 ). Scale bar=100 µm. ( b ) Effects of LOXL4 on the proliferation and migration of HUVECs. ( b1 , b2 ) There was a significant increase in the expression of LOXL4 both in intracellular and extracellular of HUVECs transfected with LOXL4 than transfected with empty plasmid. ( b3 ) Transient LOXL4 transfection increased cell growth, cell proliferation was measured with Cell Counting Kit-8 at 6, 12, 24, 72 and 96 h. ( b4 ) HUVECs migration ability was assessed by scratching assay with or without LOXL4 transfection. Representative images are shown (magnification: ×100). ( b5 ) HUVECs migration ability was also tested by Boyden Chamber assay with or without LOXL4 transfection. Representative images are shown (magnification: ×200). Each treatment was triplicate and six fields were randomly chosen from each well for counting mean number of migrated cells. Data are presented as mean±s.e.m., * P <0.05, ** P <0.01, Student's t -test. DC, dentigerous cysts; ELISA, enzyme-linked immunosorbent assay; GG, gingival primary stromal fibroblast cells; HUVEC, human umbilical vein endothelial cell; KCOT, keratocystic odontogenic tumors; LOXL4, lysyl oxidase-like 4; OD, optical density; Ctl, control.

Article Snippet: Human LOXL4 cDNA was purchased from Sino Biological and cloned in pCMV/hygro (CV004; Sino Biological, Beijng, China) for transient overexpression with a DNA Ligation Kit (2011A; TAKARA, Kyoto, Japan).

Techniques: Expressing, Immunohistochemical staining, Migration, Transfection, Plasmid Preparation, Cell Counting, Boyden Chamber Assay, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Oral Science

Article Title: Identification of the involvement of LOXL4 in generation of keratocystic odontogenic tumors by RNA-Seq analysis

doi: 10.1038/ijos.2013.96

Figure Lengend Snippet: Microvessel density and LOXL4 in keratocystic odontogenic tumors, dentigerous cysts and gingival tissues

Article Snippet: Human LOXL4 cDNA was purchased from Sino Biological and cloned in pCMV/hygro (CV004; Sino Biological, Beijng, China) for transient overexpression with a DNA Ligation Kit (2011A; TAKARA, Kyoto, Japan).

Techniques: Standard Deviation