Fig. 3. Expression profiling of store-operated Ca2+ entry (SOCE)-associated genes in Caki-1 and Caki-2 cells, with differential modulation of Orai1 and -3 by 2-aminoethoxydiphenyl borinate (2-APB).
(A) Human renal cell carcinoma (RCC) cell line models were employed for clear cell RCC (ccRCC) and papillary RCC (pRCC). (B) RT-PCR, (C, D) q-PCR for the mRNA expression levels of Orai1-3 and STIM1, 2 in Caki-1 and Caki-2, respectively. (E) Representative images displaying SOCE were recorded in cells after a one-hour pre-incubation with 2-APB (50 μM) or vehicle (control, DMSO) in Caki-1 and Caki-2, respectively. (F, G) Summary of the SOCE (F) and basal calcium (G), highlighted by the blue square in panel (E). SOCE is the difference between the peak and basal Fura-2 ratio (F340/F380), and basal Ca2+ is the average Fura-2 ratio from 0 to 60 sec. n = 60–96 cells per each group. (H, J) Representative SOCE images were recorded in Caki-1 and Caki-2 transfected with control oligo or targeting siRNAs against STIMs, respectively. (I, K, and L, M) Summary of the SOCE (I, K) and basal Ca2+ (L, M) in panels (H) and (J), respectively. n = 59–369 cells per each group. Bar graphs expressed as mean ± S.E.M. analyzed with Student’s t-test (C, D, F, and G) or one-way ANOVA (I, K, L, and M). ns., not significant; DMSO, dimethyl sulfoxide; CPA, cyclopiazonic acid. *p ≤ 0.05, ***p ≤ 0.001, ****p ≤ 0.0001. All experiments were performed three times with similar results.
© Korean J Physiol Pharmacol
