Fig. 5

TRPM1 was involved in the regulation of cytosolic Ca²⁺ concentration and photoreceptor function. a Representative western blot results of cells stably expressing either an empty vector or a vector encoding 3xFLAG-tagged wild-type TRPM1 (WT), TRPM1Ile1024Phe (I1024F) or TRPM1Cys1069Tyr (C1069Y). β-Actin was used as a loading control. n = 3. b Cytosolic Ca²⁺ levels were determined by flow cytometry using the median fluorescence intensity (MFI) of the Ca²⁺ dye Fluo-8 AM in cells expressing either an empty vector or variants of TRPM1. Data are presented as the mean ± s.e.m. The P values were determined by two-tailed Student’s t-test, * P < 0.05; ** P < 0.01; *** P < 0.001. n = 3. c GSEA of the pathways enriched in TRPM1-overexpressing 661 W cells compared with control cells based on the RNA sequencing data. NES, normalized enrichment score. d mRNA levels of Arl13b, Opn1sw and Nrf2 were determined in cells expressing either an empty vector or variants of TRPM1 by RT-qPCR. mRNA levels were calculated relative to those in cells expressing an empty vector. Levels of the housekeeping gene β-Actin were used as a reference. Data are presented as the mean ± s.e.m. The P values were determined by two-tailed Student’s t-test, * P < 0.05; ** P < 0.01; *** P < 0.001. n = 3