Journal: Cell & Bioscience
Article Title: Unraveled roles of Cav1.2 in proliferation and stemness of ameloblastoma
Figure Lengend Snippet: Cav1.2-dependent Ca 2+ /NFATc1 signaling increased the proliferation of AM cells. A – D Representative CACNA1C expression in AM cells indicated by arrowhead. CACNA1C was intensely expressed in the plasma membrane in the vehicle group. And it was broadly expressed in the plasma membrane, cytoplasm and nucleus in overexpression group. CACNA1C negatively expressed in knockdown group compared to the scramble. E – H The nuclear translocation of NFATc1 was observed in the overexpression group compared to the vehicle. NFATc1 was negatively expressed in the knockdown group compared to the scramble. I – L Representative confocal images of Ki67 staining of AM cells among the vehicle, overexpression, scramble, and siRNA groups. Nuclear were stained with TO-PRO-3 (TP3). M Percentages of CACNA1C + and NFATc1 + cells were quantified from immunostained images (N = 5 per group, biological replication). N Relative mRNA expression of CACNA1C , MKI67 , and NFATC1 between vehicle and CACNA1C overexpression, or scramble and siRNA group. O Western blot assay of AM cells with CACNA1C, PCNA, Cyclin D1, and GAPDH antibodies. P Western blot assay of nucleus and cytoplasm fractionated AM cells with NFATc1, Histone H3, and GAPDH antibodies. Scale bar: A–D, 50 μm; E–L, 100 μm. Quantitative data are presented as the mean ± SD. ** p
Article Snippet: Rabbit anti-CACNA1C (1:200, Alomone Labs, ACC-003), mouse anti-CK14 (1:500, Abcam, ab7800), mouse anti-CK10 (1:500, Invitrogen, MA5-13705), mouse anti-E-cadherin (1:500, BD Biosciences, AF748), rabbit anti-MMP-9 (1:200, Merck, AB19016), rabbit anti-LGR5 (1:200, Abcam, ab75732), mouse anti-Ki67 (1:200, Abcam, ab16667), mouse anti-PCNA (1:500, Abcam, ab29), mouse anti-NFATc1 (1:200, Santa Cruz, SC-7294), and mouse anti-β-catenin (1:500, Santa Cruz, SC-7963).
Techniques: Expressing, Over Expression, Translocation Assay, Staining, Western Blot