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. 2018 Nov 15;13(11):e0207533. doi: 10.1371/journal.pone.0207533

Fig 7. Gga1 knockdown impaired the expression of the insulin receptor.

Fig 7

(A) Total proteins (Total) and the cell surface proteins (Surface) purified using surface biotinylation from growing (day 0) and differentiated (day 4) cells were subjected to immunoblotting for golgi associated, gamma adaptin ear containing, ARF binding protein 1 (GGA1), the insulin receptor (IR), and the insulin-like growth factor 1 receptor (IGF1R). b-actin was used as the loading control. (B) Quantitative real-time PCR (qPCR) for Insr (encoding IR) was carried out. (C) Lysosomal inhibitors suppressed the degradation of the IR in Gga1kd cells. Myotubes differentiated for 4 days were treated with lysosomal inhibitors (L.I.) or proteasome inhibitor (P.I.) for 18 hours. The total lysates were subjected to immunoblotting for the IR. The signals for the ab precursor (D) and b-chain (E) of the IR were quantified and normalized by the b-actin signal. Error bars indicate the SD (n = 3). *. P < 0.05 by Welch’s t-test.