Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: An amino acid transporter subunit as an antibody–drug conjugate target in colorectal cancer

doi: 10.1186/s13046-023-02784-0

Figure Lengend Snippet: The anti-CD98hc-DM1 has anti-proliferative effect on tumoral patient-derived organoids. A Levels of expression of CD98hc in normal and tumoral PDOs. Levels of expression of CD98hc and CA2 (normal colon marker) were analyzed by Western blot with the anti-CD98hc V509 antibody. GAPDH was used as a loading control. B Immunofluorescence detection of CD98hc in tumoral PDOs. Tumoral PDOs were incubated with 10 nM of the anti-CD98hc ECTO antibody for 1.5 h and the subcellular distribution of CD98hc was analyzed by immunofluorescence. Scale bar = 50 µm. The drawing on the left represents an organoid and the red dashed lines indicate the organoid regions where images were taken. C Evaluation of anti-CD98hc ADC on the cell viability of different tumoral PDOs. The tumoral PDOs were treated with 10 nM of anti-CD98hc-DM1 for 4 days and the cell viability was analyzed as indicated in the methods sections. Results are shown as the fold-change respect to untreated organoids ± SD of triplicates of an experiment repeated twice. D Dose–response analyses of the effect of anti-CD98hc-DM1 on cell viability of paired normal and tumor PDO (#175), analyzed at four days. Results are shown as the fold-change respect to the untreated organoids ± SD of triplicates of an experiment repeated twice. P values were calculated using Student t test (two-sided). E Immunofluorescence analyses of the effect of the anti-CD98hc or anti-CD98hc-DM1 on a tumoral PDO. Organoids were treated with anti-CD98hc or anti-CD98hc-DM1 at 10 nM for 4 days and the effect on cell morphology was evaluated by inmunofluorecescence. CD98hc is stained in green, nuclei are stained in blue. Scale bars = 25 µm. Arrows indicate nuclear fragmentation

Article Snippet: Antibody against CA2 (reference CSB-PA004370GA01HU, WB: 1:1,000), used in Western blot, was from Cusabio (Houston, TX, USA).

Techniques: Derivative Assay, Expressing, Marker, Western Blot, Control, Immunofluorescence, Incubation, Staining

Journal: Archives of Toxicology

Article Title: Thallium reabsorption via NKCC2 causes severe acute kidney injury with outer medulla-specific calcium crystal casts in rats

doi: 10.1007/s00204-024-03868-2

Figure Lengend Snippet: Downregulated expression of kidney transporter and channel genes and Ca 2+ -ATPase 5 days after Tl administration at 30 mg/kg. Quantitative real-time PCR analysis of genes encoding mTAL-related kidney transporters and channels ( a ). GAPDH, glyceraldehyde-3-phosphate dehydrogenase; Kir 1.1, potassium inwardly rectifying channel subfamily J member 1; Kir 4.1, potassium inwardly rectifying channel subfamily J member 10; NaK-ATP, Na + /K + transporting ATPase alpha 1 subunit; NKCC2, Na–K–Cl cotransporter 2. GAPDH levels served as an internal control. * P < 0.05; ** P < 0.01; *** P < 0.001. Immunohistochemistry of Ca 2+ -ATPase in the outer and inner medullae of rats in the control and Tl-loaded groups ( b ). Ca 2+ -ATPase expression is decreased in the mTAL obstructed by calcium deposits (*)

Article Snippet: The sections were incubated with a 1:100 dilution of rabbit polyclonal anti-Ca 2+ -ATPase antibody (Cosmo Bio Co. Ltd., Tokyo, Japan), followed by a 1:400 dilution of horseradish peroxidase (HRP)-labeled anti-rabbit Ig antibody (Dako, Glostrup, Denmark).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control, Immunohistochemistry