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: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

Article Title: DLX3-Dependent Regulation of Ion Transporters and Carbonic Anhydrases Is Crucial for Enamel Mineralization

doi: 10.1002/jbmr.3022

Figure Lengend Snippet: Altered expression of carbonic anhydrases in the enamel organ from Dlx3K14–cKO mice. (A) Heat map showing log(RPKM) levels of carbonic anhydrases in enamel organs from Dlx3WT and Dlx3K14–cKO mice. Fold-change and significance are indicated. (B) Western blot analysis of DLX3, CA2, CA6, and actin protein levels in the enamel organ (E.O.) and salivary gland (S.G.) of Dlx3WT and Dlx3K14–cKO mice at P10. (C) Immunohistochemical detection of CA2 and CA6 (brown precipitate) in mandibular incisor longitudinal sections from Dlx3WT and Dlx3K14–cKO mice at P15. Secretory (s), transition (t), and maturation (m) stages are indicated. Scale bar = 100 μm. (D) Measurement of carbonic anhydrase activity in extracts from E.O. and S.G. from Dlx3WT and Dlx3K14–cKO mice at P10. (E) ChIP-seq analysis of DLX3 and H3K4me3 DNA binding domains in enamel organ dissected from rat mandibular incisors (continuously growing), showing DLX3 binding near the proximal promoter (arrowhead) of Car2, Car3, Car6, Car12, and Car13, but not Car1. E.O. = enamel organ; S.G. = salivary gland.

Article Snippet: Primary antibodies used: rabbit anti-DLX3 (Abcam, Cambridge, MA, USA), rabbit anti-AMELX (Abcam), rabbit anti-CA2 (GeneTex Inc., Irvine, CA, USA), rabbit anti-CA6 (Abcam).

Techniques: Expressing, Western Blot, Immunohistochemical staining, Activity Assay, ChIP-sequencing, Binding Assay

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