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. 2019 Jan 24;20(3):503. doi: 10.3390/ijms20030503

Figure 3.

Figure 3

The influence of mechanical loading on gene and protein expression of the sensory neuropeptides SP and αCGRP in RAW264.7 cells. (A,B) Relative gene expression of SP (A) and alterations of SP expression by stimulation with SP and αCGRP (B) after 4-h loading per day on two consecutive days in relation to unloaded cells (calibrator, RQ = 1) was analyzed using quantitative real-time PCR; (D) αCGRP gene expression is depicted as ΔCt of unloaded and loaded cells after 4 h/day stretching on 2 consecutive days. Normalizer: GAPDH. One sample t-test * p < 0.05, ** p < 0.01. SP, αCGRP expression n = 20, SP expression after stimulation with SP/αCGRP n = 5; (C,E) Protein concentration of SP (C) and αCGRP (E) was determined in the cell culture supernatants after 4 h/day loading on 1, 2 or 3 days using ELISA. One sample t-test * p < 0.05. n = 7–8. SP—substance P, αCGRP—alpha-calcitonin gene-related peptide, RQ—relative quantification, PCR—polymerase chain reaction, GAPDH—glyceraldehyde 3-phosphate dehydrogenase.