Journal: Cell metabolism

Article Title: Carbon source availability drives nutrient utilization in CD8 + T cells

doi: 10.1016/j.cmet.2022.07.012

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-human/mouse beta-actin , Cell Signaling , #4967.

Techniques: Luciferase, shRNA, Plasmid Preparation, Recombinant, Staining, Cell Isolation, Selection, Protease Inhibitor, Software

Journal: PLoS ONE

Article Title: 5-HT1A/1B Receptors as Targets for Optimizing Pigmentary Responses in C57BL/6 Mouse Skin to Stress

doi: 10.1371/journal.pone.0089663

Figure Lengend Snippet: A: mRNA expression analysis of MITF, TYR, TRP1 and TRP2 in 5-HT-treated HFs for 7 days. B: Quantitative results for RT-PCR amplification of 5-HTRs, 1A, 1B 1D, 2A, 5A, 5B and 7 following 5-HT-treated concentration at 10 µM, 100 µM and 1000 µM. Quantities were normalized to endogenous β-actin expression. Data are means ± SD (n = 3, *P <0.05 and **P <0.01 vs control group).

Article Snippet: The membrane was blocked with 5% non-fat dry milk in TBS containing 0.05% Tween-20 (TBS-T) for 1 h and incubated with goat polyclonal antibodies against TYR (Product number SC7833), TRP1 (Product number SC10443), rabbit polyclonal antibodies against TRP2 (Product number AB74073, 1∶1000, Abcam, Cambridge, UK), mouse polyclonal antibodies against β-actin (Product number CST3700, 1∶1000, Cell Signaling Technology Inc., MA, USA).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Amplification, Concentration Assay

Journal: Iranian Journal of Pharmaceutical Research : IJPR

Article Title: Suvorexant, a Dual Orexin Receptor Antagonist, Protected Seizure through Interaction with GABA A and Glutamate Receptors

doi: 10.22037/ijpr.2019.14688.12584

Figure Lengend Snippet: Effect of suvorexant on the protein level of NMDAR in the mice hippocampus. Suvorexant was administrated for seven days. After seizure induction by PTZ, hippocampus was dissected. Representative western blots showing specific bands for NMDAR and β-actin as an internal control. The equal amounts of protein sample (50 µg) obtained from whole hippocampus homogenate were applied in each lane. These bands are representative of four separate experiments. (B) Densitometric data of protein analysis. Data are expressed as the mean ± SEM. *** p < 0.001 vs PTZ+ normal saline. NS (normal saline), SUV (suvorexant), PTZ (pentylenetetrazol).

Article Snippet: Next, the blots were probed at room temperature with specific antibodies for 2 h. The membranes were washed three times for 5 min and incubated with HRP-conjugated secondary antibodies for 1–2 h. The primary antibodies were mouse polyclonal anti-serum against AMPA (GluR 2/3/4) (Cell signaling #2460), mouse polyclonal anti-serum against NMDA receptor (Cell signaling #4205) and mouse polyclonal anti-serum against β-actin (Cell signaling #3700).

Techniques: Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Glycosylphosphatidylinositol Mannosyltransferase Ⅰ Protects Chinese Giant Salamander, Andrias davidianus, against Iridovirus

doi: 10.3390/ijms23169009

Figure Lengend Snippet: The mRNA transcription levels of AdGPI-MT-I in six healthy Chinese giant salamander tissues including muscle, liver, spleen, thymus, kidney, heart, and intestine. The mRNA expression of AdGPI-MT-I was detected by quantitative real-time PCR (qRT-PCR), which was normalized to β-actin. The expression level in muscle was set as 1. Data are the means of three independent assays and are presented as means ± SD.

Article Snippet: Meanwhile, the expression of β-actin was detected by western blot with anti-β-actin mouse polyclonal antibodies (#12262, CST, Danvers, MA, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Glycosylphosphatidylinositol Mannosyltransferase Ⅰ Protects Chinese Giant Salamander, Andrias davidianus, against Iridovirus

doi: 10.3390/ijms23169009

Figure Lengend Snippet: Expressions of AdGPI-MT-I in the spleen ( A ), kidney ( B ), and GSM cells ( C ) at indicated times post GSIV infection. The mRNA expression level of AdGPI-MT-I was determined by quantitative real-time PCR (qRT-PCR), with six salamanders included in each time period. For the convenience of comparison, the expression level in the control at 0 h normalized to β-actin was set as 1. Error bars indicate the mean ± SD ( n = 3). ** p < 0.01, * p < 0.05. Six salamanders were included in each time period.

Article Snippet: Meanwhile, the expression of β-actin was detected by western blot with anti-β-actin mouse polyclonal antibodies (#12262, CST, Danvers, MA, USA).

Techniques: Infection, Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Glycosylphosphatidylinositol Mannosyltransferase Ⅰ Protects Chinese Giant Salamander, Andrias davidianus, against Iridovirus

doi: 10.3390/ijms23169009

Figure Lengend Snippet: ( A ) Western blot was used to verify the overexpression of the AdGPI-MT-I protein in GSM cells. GSM cells were transfected with pmCherryN1 vector or pmCherryN1-AdGPI-MT-I plasmid DNA for 48 h and then harvested for western blot. β-actin was used as an internal reference. ( B ) Confocal microscopy was used to observe the subcellular localization of the AdGPI-MT-I protein in GSM cells (scale bar, 20 μm). Nuclei were stained with DAPI. The nucleus and cytoplasm are marked with the numbers 1 and 2, respectively.

Article Snippet: Meanwhile, the expression of β-actin was detected by western blot with anti-β-actin mouse polyclonal antibodies (#12262, CST, Danvers, MA, USA).

Techniques: Western Blot, Over Expression, Transfection, Plasmid Preparation, Confocal Microscopy, Staining

Journal: International Journal of Molecular Sciences

Article Title: Glycosylphosphatidylinositol Mannosyltransferase Ⅰ Protects Chinese Giant Salamander, Andrias davidianus, against Iridovirus

doi: 10.3390/ijms23169009

Figure Lengend Snippet: ( A ) Western blot analysis of MCP protein expression and grayscale quantitative analysis. Western blot was used to detect protein synthesis after GSIV infection at different times, and mouse anti-MCP monoclonal antibody and anti-β-actin monoclonal antibodies were used as primary antibodies, respectively. HRP-tagged antimouse IgG was used as a secondary antibody. β-actin was used as internal control. The gray values of GSIV-MCP and β-actin protein bands were quantitatively analyzed by ImageJ software. ( B ) Digital droplet PCR (ddPCR) was used to detect the MCP gene copy number of GSIV in GSM cells. After transfection of pmCherryN1-AdGPI-MT-I plasmid with GSM cells for 48 h, the copy number of the MCP gene at 0, 12, 24, 48, and 72 h after GSIV infection was detected by ddPCR. Error bars indicate the mean ± SD ( n = 3). The asterisks indicate significant difference (* p < 0.05) between treated and control groups.

Article Snippet: Meanwhile, the expression of β-actin was detected by western blot with anti-β-actin mouse polyclonal antibodies (#12262, CST, Danvers, MA, USA).

Techniques: Western Blot, Expressing, Infection, Software, Transfection, Plasmid Preparation

Journal: Bioengineered

Article Title: P75 neurotrophin receptor positively regulates the odontogenic/osteogenic differentiation of ectomesenchymal stem cells via nuclear factor kappa-B signaling pathway

doi: 10.1080/21655979.2022.2063495

Figure Lengend Snippet: Odonto/osteogenic differentiation in NF-κB-inhibited EMSCs. (a) Under induction with dental epithelium conditioned medium for 7 days, ALP staining was used to detect the potential of odonto/osteogenic differentiation in p75NTR +/+ EMSCs group and p75NTR +/+ EMSCs-inhibitor group. ALP activities of p75NTR +/+ EMSCs-inhibitor was significantly lower compared with p75NTR +/+ EMSCs. (b) Alizarin red staining was used to detect the potential of odonto/osteogenic differentiation in p75NTR +/+ EMSCs group and p75NTR +/+ EMSCs-inhibitor group. Calcium quantification illustrated the higher calcium deposition in p75NTR +/+ EMSCs compared with p75NTR +/+ EMSCs-inhibitor group. (c) RT-PCR for the detection of DSPP, Runx2, DMP1, ALP, OCN and OSX of EMSCs, respectively, p75NTR +/+ EMSCs, p75NTR −/− EMSCs, and p75NTR +/+ EMSCs + inhibitor groups at day 7. (d) After 7 days of induction, the expression levels of P65, p-P65 and DSPP were detected by western blot, β-Actin used as the reference gene. Semiquantitative analysis demonstrated that the expression odonto /osteogenic marker (DSPP) was significantly downregulated in NF-κB pathway-inhibited EMSCs than those in control group at day 7. The data are presented as mean ± SD, n = 3, *P < 0.05, **P < .01, ***P < .001. Scale bar, 100 μm.

Article Snippet: The primary antibodies used in this experiment were as follows: rabbit anti- polyclonal RUNX2 (1: 1000, Abcam,USA), rabbit anti- polyclonal DSPP (1: 1000, Absin,USA), rabbit anti- polyclonal P- NF-κB (1: 1000,CST,USA), rabbit anti- polyclonal NF-κB (1:1000, CST,USA), and mouse anti- polyclonal β-ACTIN (1: 8000, CST,USA), IRDye® 800CW Goat anti-Rabbit(1:5000,LI-COR,USA) and IRDye® 680RD Goat anti-Mouse(1:5000,LI-COR,USA).The membranes were visualized and scanned by Odyssey Clx(LI-COR,USA).

Techniques: Staining, Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot, Marker