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. 2019 Sep 13;33(11):12853–12872. doi: 10.1096/fj.201900057R

Figure 4.

Figure 4

Pharmacological evidence implicating TRPC1 in magnetoreception. A) Effect of 2-APB (10 μM; hatched) on PEMF-induced proliferation at 24 h (dark blue) and 48 h (black) after plating; n = 11 (24 h) and n = 7 (48 h). B) Effect of SKF-96365 (50 µM) on PEMF-induced proliferation; n = 5. C) Levels of [Ca2+]i following 10 min exposure to 0 (red) or 1 mT (blue) PEMFs in the absence (solid) or presence (hatched) of 10 mM MgCl2 (hexahydrate) added to the culture medium immediately before exposure; n = 3 ± sem). D) PEMF-modulated proliferation (Prolif; 1.5/0 mT) in response to no antibiotics (solid), penicillin/streptomycin (PS) (1%, hatched), streptomycin (Strepto; 100 mg/L), neomycin (Neo; 50 mg/L), or gentamycin (Genta; 50 mg/L) added to the culture medium at time of PEMF exposure; n = 4. Inset: effect of 10 min PS application and immediate removal 2 h before PEMF exposure; n = 3. E, F) PS (1%) (E) and Neo (50 mg/L) (F) prevented myotube formation following single PEMF exposure. G–L) Myotube formation without (G–I) and with (J–L) PS (1%) as indicated. Scale bar, 100 μm. M) PS and 2-APB attenuate 2-APB–sensitive PEMF-induced calcium entry; n = 2 × 8 wells. N, O) Myotube size distribution without (N) and with (O) CsA; correspondent to Fig. 2I. The red dashed lines indicate the previous 0 mT level. Unless otherwise stated, all data are generated from the means of independent experiments ± sd each pertaining to minimally the means of biological triplicates. All PEMF exposures were conducted for 10 min applied 24 h postplating, unless otherwise stated (48 h; A); 24 h post-PEMF exposure myoblasts were either counted (B–D) or allowed to differentiate (E, F) for 6 d. **P < 0.01 (with regard to 0 and 1.5 mT, black and red asterisks, respectively). All drugs were applied transiently to coincide with PEMF exposure and to avoid non-TRP channel–related tertiary effects. Specifically, 2-APB, SKF-96365, and antibiotics were added to culture medium before PEMF exposure and then replaced with age-matched control medium from sister cultures summarily thereafter, with the exception of D (inset), wherein PS (hatched) was added and removed before PEMF exposure.