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Fig. 4 | Cell Communication and Signaling

Fig. 4

From: Peptide targeting of lysophosphatidylinositol-sensing GPR55 for osteoclastogenesis tuning

Fig. 4

LPI induces increases in intracellular Ca2+ through an endogenous GPR55 in RAW264.7 cells. a Gpr55 mRNA levels of RAW264.7 cells interfered with non-targeting (si-NT) or Gpr55-specific (si-GPR55) siRNAs were quantified by real-time PCR and normalised using β2-microglobulin expression, as the housekeeping gene. Data are means ± SEM from seven independent experiments. b, c Interfered RAW264.7 cells were serum deprived for 1 h, loaded with Fluo4-NW, and then stimulated without or with 10 µM 16:0 LPI. Changes in fluorescence of each well of a 96-well plate were recorded, and the intracellular Ca2+ concentrations were calculated (see “Materials and methods” section). b Comparison between si-NT-and si-GPR55-responses to 10 µM 16:0 LPI, as means of six independent experiments performed in quadruplicates. Amplitude (height of the Ca2+ peak measured at the apex) and duration (length of the peak measured at the bases) of the responses were measured. c Data are means ± SE of quadruplicates within a representative experiment. *p < 0.05; ***p < 0.005 versus si-NT (Student’s t-tests)

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