rabbit polyclonal antibody against total akt  (Cell Signaling Technology Inc)

Journal: Scientific Reports

Article Title: The effects of artocarpin on wound healing: in vitro and in vivo studies

doi: 10.1038/s41598-017-15876-7

Figure Lengend Snippet: ARTO enhances human fibroblast proliferation and migration through the P38 or JNK signaling pathway. ( a ) Immunohistochemistry was performed to identify vimentin and PCNA (arrows) in wounds on day 15. The cells were pre-treated with MK2006 (MK), PD98059 (PD), SB203580 (SB), or SP600125 (SP) for 1 h and then incubated with ARTO for 24 h. Crystal violet staining ( b ) and a BrdU incorporation assays ( c ) were used to measure GM05386 cell number and proliferation. ( d ) Cell migration was examined via wound healing assays, in which GM05386 cells were wounded by scratch injury (black lines). The wound closure and migration rates were determined. The levels of Akt ( e ), ERK ( f ), P38 ( g ), and JNK ( h ) were determined by western blot analysis. ( i ) There was co-localization between vimentin and phosphorylated P38 or JNK in the wounds on day 15. The data are shown as the means ± S.D. N = 3–6 group and * P < 0.05.

Article Snippet: Polyclonal rabbit IgG antibodies against human GAPDH, β-actin, phospho-/total-P38, phospho-/total-ERK1/2, phospho-/total-JNK, α-actin, collagen, phospho-/total-Akt, horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG, and anti-rabbit IgG antibodies were purchased from GeneTex (Irvine, CA, USA).

Techniques: Migration, Immunohistochemistry, Incubation, Staining, BrdU Incorporation Assay, Western Blot

Journal: Scientific Reports

Article Title: The effects of artocarpin on wound healing: in vitro and in vivo studies

doi: 10.1038/s41598-017-15876-7

Figure Lengend Snippet: ARTO enhances human keratinocyte proliferation and migration through the ERK or P38 signaling pathway. ( a ) Wounded sections were stained with CK14 and PCNA (arrows). ( b ) Quantitative analysis of PCNA-positive cells. The cells were pre-treated with PD98059 (PD) or SB203580 (SB) for 1 h and then incubated with ARTO for 24 h. Crystal violet staining ( c ), western blot analysis ( d ), and BrdU incorporation assays ( e ) were used to measure keratinocyte proliferation. ( f ) Quantitative analysis of BrdU-positive cells. ( g , h ) The wound closure and migration rates were examined via wound healing assays, in which keratinocytes were wounded by scratch injury (black lines). The levels of Akt ( i ), ERK ( j ), P38 ( k ), and JNK ( l ) were determined by western blot analysis. ( m ) There was co-localization between CK14 and phosphorylated ERK or P38 in the wounds on day 7. The data are shown as the means ± S.D. N = 3–6 wounds/group and * P < 0.05.

Article Snippet: Polyclonal rabbit IgG antibodies against human GAPDH, β-actin, phospho-/total-P38, phospho-/total-ERK1/2, phospho-/total-JNK, α-actin, collagen, phospho-/total-Akt, horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG, and anti-rabbit IgG antibodies were purchased from GeneTex (Irvine, CA, USA).

Techniques: Migration, Staining, Incubation, Western Blot, BrdU Incorporation Assay

Journal: Scientific Reports

Article Title: The effects of artocarpin on wound healing: in vitro and in vivo studies

doi: 10.1038/s41598-017-15876-7

Figure Lengend Snippet: ARTO enhances angiogenesis through the Akt or P38 signaling pathway. ( a , b ) Wounded sections were stained with CD31, and a quantitative analysis of CD31 level was conducted. ( c ) Western blot analysis of the skin showed that CD31 level was increased in the ARTO-treated wounds on day 15 after wounding. ( d ) Immunohistochemistry was performed to identify CD31 and PCNA (arrows) in wounds on day 15. ( e , f ) MTT, crystal violet staining, and BrdU incorporation assays were used to measure cell viability and proliferation. The levels of Akt ( g ), ERK ( h ), P38 ( i ), and JNK ( j ) were determined by western blot analysis. HUVECs were pre-treated with Akt or MAPK inhibitors for 1 h and then incubated with ARTO for 24 h. ( k ) Crystal violet staining was performed. ( l ) Tube formation and tube length were examined by a Matrigel assay. Confocal image of a tube stained for CD31 (green) and cell nuclei (DAPI, blue). The asterisk indicates the lumen of the tube. ( m ) There was co-localization between CD31 and phosphorylated Akt or P38 in the wounds on day 15. The data are shown as the means ± S.D. N = 3–6 wounds/group and * P < 0.05.

Article Snippet: Polyclonal rabbit IgG antibodies against human GAPDH, β-actin, phospho-/total-P38, phospho-/total-ERK1/2, phospho-/total-JNK, α-actin, collagen, phospho-/total-Akt, horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG, and anti-rabbit IgG antibodies were purchased from GeneTex (Irvine, CA, USA).

Techniques: Staining, Western Blot, Immunohistochemistry, BrdU Incorporation Assay, Incubation, Matrigel Assay