mouse ab against β-actin  (MBL Life science)

Journal: Cell Journal (Yakhteh)

Article Title: Downregulation of Vascular Endothelial Growth Factor Enhances Chemosensitivity by Induction of Apoptosis in Hepatocellular Carcinoma Cells

doi:

Figure Lengend Snippet: Effects of VEGF-siRNA on VEGF mRNA expression in Hep3B cells. Cells were transfected with VEGF-siRNA and CONT-siRNA, then harvested at indicated times. Total RNA was extracted from cells at the indicated time after siRNA transfection. A. Electrophoretic profile of PCR products of the VEGF (250 bp) and β-actin (680 bp) genes. β-actin was used as a housekeeping gene control and B. Quantitative analyses of VEGF mRNA levels were measured by real-time quantitative PCR (qRT-PCR). mRNA expression of VEGF was normalized with β-actin. Each bar represents the mean value ± standard deviation (SD) of triplicate. **; P<0.01 compared to untreated cell group. VEGF-siRNA; Vascular endothelial growth factor targeted smallinterfering RNA, CONT-siRNA; Mismatched small-interfering RNA and PCR; Polymerase chain reaction.

Article Snippet: Mouse monoclonal Ab against β-actin (Abcam, Cambridge, England, UK) was used as a housekeeping gene control.

Techniques: Expressing, Transfection, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Standard Deviation, Small Interfering RNA, Polymerase Chain Reaction

Journal: Cell Journal (Yakhteh)

Article Title: Downregulation of Vascular Endothelial Growth Factor Enhances Chemosensitivity by Induction of Apoptosis in Hepatocellular Carcinoma Cells

doi:

Figure Lengend Snippet: Effects of VEGF-siRNA on VEGF production and secretion in Hep3B cells. Total cellular protein was extracted from cells at an indicated time after siRNA transfection. After electrophoresis and electrotransfer, the membranes were incubated with mouse Ab against VEGF followed by HRP-linked goat anti-mouse IgG. A. Western blot analysis of VEGF protein expression in Hep3B cells. β-actin was used as a housekeeping gene control. The size of each protein was indicated. B. VEGF downregulated Hep3B cells exhibited decreased expression of VEGF protein as confirmed by densitometric analysis and C. The cell culture supernatants were collected at different time intervals and the secreted VEGF concentrations measured by a quantitative VEGF ELISA kit. Each bar represents the mean value ± standard deviation (SD) of triplicate analyses. *; P<0.05, **; P<0.01 compared to untreated cell group, VEGF-siRNA; Vascular endothelial growth factor targeted small-interfering RNA and ELIZA; Enzyme-linked ammunosorbent assay.

Article Snippet: Mouse monoclonal Ab against β-actin (Abcam, Cambridge, England, UK) was used as a housekeeping gene control.

Techniques: Transfection, Electrophoresis, Electrotransfer, Incubation, Western Blot, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay, Standard Deviation, Small Interfering RNA

Journal: Cell Journal (Yakhteh)

Article Title: Downregulation of Vascular Endothelial Growth Factor Enhances Chemosensitivity by Induction of Apoptosis in Hepatocellular Carcinoma Cells

doi:

Figure Lengend Snippet: Effect of VEGF-siRNA and/or doxorubicin treatment on BCL-2 and SURVIVIN mRNA expression in Hep3B cells. A, C. mRNA expressions of BCL-2 and SURVIVIN in cells detected by reverse transcription polymerase chain reaction (RT-PCR) after treatment with the indicated concentration of doxorubicin for 48 hours. Electrophoretic profile of PCR products of BCL-2 (174 bp), SURVIVIN (145 bp) and β-actin (680 bp) genes. β-actin was used as a housekeeping gene control and B, D. mRNA levels of BCL-2 and SURVIVIN in cells determined by real-time quantitative PCR (qRT-PCR). mRNA expressions of these genes were normalized with β-actin. Each bar represents the mean value ± standard deviation (SD) of triplicate experiments. *; P<0.05, **; P<0.01 compared to control cell group (or medium), #; P<0.05, ##; P<0.01 compared to VEGF-siRNA treated cell group, Dox 0; 0 μg/ml (or medium) doxorubicin, Dox 1; 1 μg/ml doxorubicin, Dox 2; 2 μg/ml doxorubicin, Dox 4; 4 μg/ml doxorubicin and VEGF-siRNA; Vascular endothelial growth factor targeted small-interfering RNA

Article Snippet: Mouse monoclonal Ab against β-actin (Abcam, Cambridge, England, UK) was used as a housekeeping gene control.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Concentration Assay, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Standard Deviation, Small Interfering RNA

Journal: Cell Journal (Yakhteh)

Article Title: Downregulation of Vascular Endothelial Growth Factor Enhances Chemosensitivity by Induction of Apoptosis in Hepatocellular Carcinoma Cells

doi:

Figure Lengend Snippet: Effect of VEGF-siRNA and/or doxorubicin treatment on BCL-2 and SURVIVIN protein expression in Hep3B cells. A. Protein expressions of BCL-2 and SURVIVIN in Hep3B cells measured by Western blot analyses after treatment with the indicated concentration of doxorubicin for 48 hours. β-actin was used as a housekeeping gene control. The size of each protein was indicated and B, C. Densitometric analysis of these three proteins relative to β-actin. Each bar represents the mean value ± standard deviation (SD) of triplicate experiments. *; P<0.05, **; P<0.01 compared to control cell group (or medium), #; P<0.05 compared to VEGF-siRNA treated cell group, Dox 0; 0 μg/ml (or medium) doxorubicin, Dox 1; 1 μg/ml doxorubicin, Dox 2; 2 μg/ml doxorubicin, Dox 4: 4 μg/ml doxorubicin and VEGF-siRNA; Vascular endothelial growth factor targeted small-interfering RNA.

Article Snippet: Mouse monoclonal Ab against β-actin (Abcam, Cambridge, England, UK) was used as a housekeeping gene control.

Techniques: Expressing, Western Blot, Concentration Assay, Standard Deviation, Small Interfering RNA

Journal: Frontiers in Immunology

Article Title: The Non-Receptor Protein Tyrosine Phosphatase PTPN6 Mediates a Positive Regulatory Approach From the Interferon Regulatory Factor to the JAK/STAT Pathway in Litopenaeus vannamei

doi: 10.3389/fimmu.2022.913955

Figure Lengend Snippet: Tissue distribution and subcellular localization of LvPTPN6. (A) Expression of LvPTPN6 in L. vannamei tissues detected by qRT-PCR with EF-1α as internal control. The expression level of LvPTPN6 in pyloric cecum was set as the baseline (1.0). Each bar represents the mean ± SD ( n = 4). (B) Subcellular localization of HA-tagged LvPTPN6 was detected by confocal laser scanning microscopy analysis in S2 cells. LvPTPN6 was stained with Alexa Fluor 488 (green), the cytomembranes were visualized by β-actin stain with Alexa Flour 594 (red), and the nuclei were stained with Hoechst 33342 (blue).

Article Snippet: For nuclear translocation of L. vannamei STAT, shrimps were challenged with Poly(I:C) for 12 h as mentioned above, then hemolymph smear samples were made on siliconized slides and fixed with 4% paraformaldehyde for 10 min. For immunofluorescence assay, 4% paraformaldehyde fixed cells were infiltrated with 1% Triton X-100 for 20 min then successively incubated with rabbit Ab against HA (CST, Danvers, MA, USA) or shrimp STAT (GL Biochem, Shanghai, China) together with mouse Ab against β-actin (MBL, Tokyo, Japan) and Alexa Fluor 488-conjugated goat anti-rabbit Ab (CST, USA) together with Alexa Fluor 594-conjugated goat anti-mouse Ab (CST, USA).

Techniques: Expressing, Quantitative RT-PCR, Confocal Laser Scanning Microscopy, Staining

Journal: Frontiers in Immunology

Article Title: The Non-Receptor Protein Tyrosine Phosphatase PTPN6 Mediates a Positive Regulatory Approach From the Interferon Regulatory Factor to the JAK/STAT Pathway in Litopenaeus vannamei

doi: 10.3389/fimmu.2022.913955

Figure Lengend Snippet: Nuclear localization of STAT regulated by LvPTPN6. (A) qRT-PCR analysis of the LvPTPN6 mRNA level in hemocyte and gill at 48 h post dsRNA injection. Values in the dsRNA-GFP control group were set as the baseline (1.0). Each bar represents the mean ± SD ( n = 4), ** p < 0.01 by two-tailed unpaired Student’s t -test. (B) Western blot analysis of the protein level of STAT located in the cytoplasm and nucleus of hemocytes after dsRNA injection. (C) Immunofluorescence intensities (arbitrary units, AU) of cytoplasm- and nuclear-localized STAT in dsNRA-GFP- and dsRNA-PTPN6-treated hemocytes. Immunofluorescence intensities were calculated using JACoP with an ImageJ plugin from four randomly selected microscopic vision fields ( Supplemental Figure 3 ). * p < 0.05 by two-tailed unpaired Student’s t -test. (D) Immunofluorescent analysis of STAT in hemocytes after dsRNA injection. STAT was stained with Alexa Fluor 488 (green), the cytomembranes were visualized by β-actin stain with Alexa Flour 594 (red), and the nuclei were stained with Hoechst 33342 (blue). (E) Western blot analysis of the protein level of shrimp STAT located in the cytoplasm and nucleus of S2 cells overexpressed with LvPTPN6 or GFP (negative control). (B, E) In cytoplasm, the gray values of STAT bands were normalized to those of the cytoplasmic internal control of β-actin, and Histone H3 was detected to verify no contamination of nuclear protein. In nucleus, the gray values of STAT bans were normalized to those of the nuclear internal control of Histone H3, and β-actin was detected to verify no contamination of cytoplasmic protein. (F) Western blot analysis of the dimer and monomer levels of STAT through native PAGE after overexpressing LvPTPN6 in S2 cells. The gray values of STAT bans were normalized to those of the internal control of β-actin. (B–F) Each bar is mean ± SD of three independent quantification of the electrophoretic bands, ** p < 0.01 by two-tailed unpaired Student’s t -test.

Article Snippet: For nuclear translocation of L. vannamei STAT, shrimps were challenged with Poly(I:C) for 12 h as mentioned above, then hemolymph smear samples were made on siliconized slides and fixed with 4% paraformaldehyde for 10 min. For immunofluorescence assay, 4% paraformaldehyde fixed cells were infiltrated with 1% Triton X-100 for 20 min then successively incubated with rabbit Ab against HA (CST, Danvers, MA, USA) or shrimp STAT (GL Biochem, Shanghai, China) together with mouse Ab against β-actin (MBL, Tokyo, Japan) and Alexa Fluor 488-conjugated goat anti-rabbit Ab (CST, USA) together with Alexa Fluor 594-conjugated goat anti-mouse Ab (CST, USA).

Techniques: Quantitative RT-PCR, Injection, Two Tailed Test, Western Blot, Immunofluorescence, Staining, Negative Control, Clear Native PAGE

Journal: PLoS ONE

Article Title: Adipocyte STAT5 deficiency does not affect blood glucose homeostasis in obese mice

doi: 10.1371/journal.pone.0260501

Figure Lengend Snippet: Abundance of total STAT3 and of phosphorylated STAT3 (pSTAT3) normalised to β-actin in epidydimal fat (A) and UCP1 expression normalised to β-actin in the inguinal fat (B) from 23–24 weeks old male mice fed high-fat diet ad libitum which lack STAT5 in their adipocytes ( Stat5 Adipoq / KO) as compared to their wild type controls (WT). No significant differences WT vs Stat5 Adipoq .

Article Snippet: For the detection of STAT3 and phospho-STAT3 Rabbit primary Ab against STAT3 and phospho-STAT3 Tyr705 (#4904 and #9145, Cell Signaling Technology, Danvers, MA, USA) were used at a dilution of 1:1,000 in TBST, and Mouse primary Ab against β-actin as house-keeping protein (#ab8226, Abcam, Cambridge, UK) were used at a dilution of 1:5,000 in TBST.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Adipocyte STAT5 deficiency does not affect blood glucose homeostasis in obese mice

doi: 10.1371/journal.pone.0260501

Figure Lengend Snippet: Abundance of total STAT3 and of phosphorylated STAT3 (pSTAT3) normalised to β-actin in epidydimal fat (A) and UCP1 expression normalised to β-actin in the inguinal fat (B) from 23–24 weeks old male mice fed high-fat diet ad libitum which lack STAT5 in their adipocytes ( Stat5 Adipoq / KO) as compared to their wild type controls (WT). No significant differences WT vs Stat5 Adipoq .

Article Snippet: For the detection of STAT3 and phospho-STAT3 Rabbit primary Ab against STAT3 and phospho-STAT3 Tyr705 (#4904 and #9145, Cell Signaling Technology, Danvers, MA, USA) were used at a dilution of 1:1,000 in TBST, and Mouse primary Ab against β-actin as house-keeping protein (#ab8226, Abcam, Cambridge, UK) were used at a dilution of 1:5,000 in TBST.

Techniques: Expressing