primary antibody against canx  (Proteintech)

Journal: Journal of Cancer

Article Title: n6-methyladenosine-modified circular RNA family with sequence similarity 126, member A affects cholesterol synthesis and malignant progression of prostate cancer cells by targeting microRNA-505-3p to mediate calnexin

doi: 10.7150/jca.89135

Figure Lengend Snippet: Sequence of all genes.

Article Snippet: Primary antibodies against CANX (1:1000, ab22595, Abcam), insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1, 1:1000, ab82968, Abcam), Vascular endothelial growth factor (VEGF, 1:1000, sc-7269, Santa Cruz Biotechnology), programmed death-ligand 1 (PD-L1, 1:1000, 13684, Cell Signaling Technology), and GAPDH (1:1000; ab37168, Abcam) were incubated with the membrane at 4°C overnight, and horseradish peroxidase-linked secondary antibody (1:500; SC-2054; Santa Cruz Biotechnology) was added for 2 h at room temperature.

Techniques: Sequencing

Journal: Journal of Cancer

Article Title: n6-methyladenosine-modified circular RNA family with sequence similarity 126, member A affects cholesterol synthesis and malignant progression of prostate cancer cells by targeting microRNA-505-3p to mediate calnexin

doi: 10.7150/jca.89135

Figure Lengend Snippet: circFAM126A sponges miR-505-3p to upregulate CANX. A: Bioinformatics websites PITA, microT, TargetScan, PicTar, and the miRanda database were utilized to predict potential downstream mRNA targets of miR-505-3p. B and C: The impact of knocking down or overexpressing miR-505-3p on the mRNA and protein expression of CANX was examined using RT-qPCR and Western blot. D: Dual-luciferase reporter assay was conducted to investigate the target relationship between miR-505-3p and CANX. E: The bioinformatics website starbase was employed to predict potential binding sites between CANX and miR-505-3p. F: Biotinylated pull-down assay was used to analyze the targeting relationship between miR-505-3p and CANX. G and H: RT-qPCR and Western blot assays were carried out to measure the mRNA and protein expression of CANX in PCa tissues and adjacent normal tissues. I and J: Pearson correlation analysis was performed to assess the relationship among CircFAM126A, miR-505-3p, and CANX in cancer tissues. K and L: The effect of knocking down CircFAM126A on the mRNA and protein levels of CANX was detected using RT-qPCR and Western blot. Data are presented as mean ± SD (n = 3), with * P < 0.05 indicating statistical significance.

Article Snippet: Primary antibodies against CANX (1:1000, ab22595, Abcam), insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1, 1:1000, ab82968, Abcam), Vascular endothelial growth factor (VEGF, 1:1000, sc-7269, Santa Cruz Biotechnology), programmed death-ligand 1 (PD-L1, 1:1000, 13684, Cell Signaling Technology), and GAPDH (1:1000; ab37168, Abcam) were incubated with the membrane at 4°C overnight, and horseradish peroxidase-linked secondary antibody (1:500; SC-2054; Santa Cruz Biotechnology) was added for 2 h at room temperature.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Luciferase, Reporter Assay, Binding Assay, Pull Down Assay

Journal: Journal of Cancer

Article Title: n6-methyladenosine-modified circular RNA family with sequence similarity 126, member A affects cholesterol synthesis and malignant progression of prostate cancer cells by targeting microRNA-505-3p to mediate calnexin

doi: 10.7150/jca.89135

Figure Lengend Snippet: CANX overexpression re-activates cancer malignancy after inhibition of circFAM126A or overexpression of miR-505-3p. Sh-circFAM126A and miR-505-3p mimic were co-transfected into PC-3 and DU145 cells. A: The impact of co-transfection on miR-505-3p levels in cells was measured using RT-qPCR. B: Cell proliferation rate was assessed through the CCK-8 assay. C: Colony formation assay was conducted to evaluate the cell proliferation capacity. D: EdU incorporation was used to detect the rate of Edu-positive cells. E: Flow cytometry was performed to examine cell apoptosis. F and G: Transwell assays were utilized to assess the migration and invasion capabilities of the cells. H and I: Commercial kits were employed to measure the levels of TG and cholesterol in the cells. J and K: mRNA levels of LUR1 and SREBPs in the cells were detected by RT-qPCR. Data are presented as mean ± SD (n = 3), with * P < 0.05 indicating statistical significance.

Article Snippet: Primary antibodies against CANX (1:1000, ab22595, Abcam), insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1, 1:1000, ab82968, Abcam), Vascular endothelial growth factor (VEGF, 1:1000, sc-7269, Santa Cruz Biotechnology), programmed death-ligand 1 (PD-L1, 1:1000, 13684, Cell Signaling Technology), and GAPDH (1:1000; ab37168, Abcam) were incubated with the membrane at 4°C overnight, and horseradish peroxidase-linked secondary antibody (1:500; SC-2054; Santa Cruz Biotechnology) was added for 2 h at room temperature.

Techniques: Over Expression, Inhibition, Transfection, Cotransfection, Quantitative RT-PCR, CCK-8 Assay, Colony Assay, Flow Cytometry, Migration