Figure 5

Helminth factors impair store-mediated TRPC1-STIM1 and ORAI1-STIM1 channel assembly. (A) Immunoprecipitation using anti-STIM1 antibodies, of equal amount of lysates from control nonstimulated (NS) cells, treated with or without lipopolysaccharide (LPS), or cells treated with helminth soluble factors (HSFs) (25 μg/ml for 20 minutes) and stimulated with or without LPS. Immunoblotting was performed using anti-TRPC1, anti-ORAI1 and anti-STIM1 antibodies at 1:1,000 dilution each. ORAI1, ORAI Ca²⁺ release-activated Ca²⁺ modulator 1; STIM1, Stromal interaction molecule 1; TRPC1, Transient receptor potential channel 1. (B) Bar graphs show the densitometric values of TRPC1 and ORAI1. Values are expressed as mean ± SE. **P < 0.05 versus HSF–untreated, LPS-stimulated cells. (C) Immunoprecipitation using anti-STIM1 antibodies in amounts equal to lysates from control NS cells, treated with or without thapsigargin (Tg), or cells treated with HSF (25 μg/ml for 20 minutes) and stimulated with or without Tg and immunoblotted using the respective antibodies. (D) The average pixel intensity of the respective protein bands from three independent experiments was measured using Quantity One 1-D version 4.6.7 software (Bio-Rad Laboratories). Asterisks indicate significance (*P ≤ 0.05, **P ≤ 0.01).