human lymph node tissue  (Santa Cruz Biotechnology)

Journal: Pharmaceuticals

Article Title: Increased MAGE-C Family Gene Expression Levels as a Biomarker of Colon Cancer Through the Demethylation Mechanism

doi: 10.3390/ph17111447

Figure Lengend Snippet: Identification of MAGE-C gene transcripts in tissue specimens collected from male patients with adjacent NC and CC tissues. ( A ) The RT-PCR results for the MAGE-C family genes are shown on agarose gel pictures. The cDNAs were created from the total RNA taken from 20 tissue samples, 20 of which were NC and 20 of which were CC. To ensure the integrity of the cDNA samples, the expression of the ACTB gene was evaluated, yielding a band of approximately 553 bp. The primers for each gene were validated using cDNA obtained from testicular tissue. The official names of the individual genes along with their expected product sizes are shown to the left of the gel images. ( B ) qRT-PCR analysis of MAGE-C family gene expression was measured in NC and CC samples. This qRT-PCR quantified the mRNA expression of these genes in CC samples relative to their respective NC samples, with results normalized to GAPDH mRNA levels. The error bars in the figure indicate the standard error of the mean from three separate qRT-PCR experiments for each gene. Significance is indicated by ** p ≤ 0.01, *** p ≤ 0.001, and **** p ≤ 0.0001. Abbreviations: NC (normal colon); CC (colon cancer).

Article Snippet: Additionally, to further validate the specificity of each MAGE-C primer, RT-PCR was conducted using cDNA synthesized from RNA extracted from human testis tissue (AM7972; Thermo Fisher Scientific) known to express MAGE-C genes.

Techniques: Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Expressing, Quantitative RT-PCR, Gene Expression

Journal: Platelets

Article Title: The bile acid receptor TGR5 inhibits platelet activation and thrombus formation.

doi: 10.1080/09537104.2024.2322733

Figure Lengend Snippet: Figure 4. Platelets express the bile acid receptor TGR5. Human platelets lysates (40×106) were analyzed regarding their protein expression of TGR5 (A) with and without CRP stimulation. Lysates of human gallbladder tissue and TGR5 transfected HEK293 cells served as positive control. (B) Quantification of TGR5 expression in human platelets lysates (n = 3). (C) Human platelets were allowed to adhere on a fibrinogen matrix and were stained for TGR5 and Actin filaments (n = 3). (D) Analysis of cDNA isolated from murine platelets were analyzed regarding the expression of TGR5 mRNA (n = 3). (E) Murine platelets were allowed to adhere on a fibrinogen matrix and were stained for TGR5 and actin filaments (n = 3). (F) Murine platelets of wildtype and TGR5-/- mice were pre-incubated with TCDC, TLC or DMSO control for 1 h. After preincubation, platelets were allowed to adhere on a fibrinogen matrix for 60 min and total adhesion was recorded (n = 3–6). Surface coverage and number of adherent cells (E) was determined using Fiji software. Statistical significance was determined by two-way ANOVA with Tukey’s multiple comparisons test. *p < .05, *p < .001. Data shown as mean + SEM.

Article Snippet: For immunoblotting targeting human TGR5, an antiTGR5 antiserum (M39) was raised against amino acids 298–318 of human TGR5 (NP_733800) and characterized using TGR5YFP transfected HEK293 cells and human gallbladder tissue as previously described.19 Glyceraldehyde 3-phosphate dehydrogenase (GAPDH, # Sc -32233, Santa Cruz Biotechnology, 1:10000) and horseradish peroxidase-conjugated anti-mouse secondary antibody (#P0447, DAKO, 1:5000) served as loading control.

Techniques: Expressing, Transfection, Positive Control, Staining, Isolation, Incubation, Control, Software