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Figure 9 | BMC Molecular Biology

Figure 9

From: Single-cell qPCR on dispersed primary pituitary cells -an optimized protocol

Figure 9

Single-cell qPCR on primary culture from cod pituitary using specific primers for follicle-stimulating hormone beta subunit ( FSHβ ), luteinizing hormone beta subunit ( LHβ ), and elongation factor 1 α ( EF1α ). Melting curve plotted as the negative change in fluorescence per time as a function of temperature. The different melting peaks for each qPCR product in the melting curve analysis indicates the specific phenotype of the cell. EF1α is only used as a positive control. Using a silanized patch glass, and β-escin as perforating agent prevented false positives and allowed reliable detection and to discriminate between FSH-, LH-, or non gonadotrope-producing cell. A cell positive for FSHβ and EF1α and negative for LHβ (black line without any specific peak). B cell positive for LHβ and EF1α and negative for FSHβ (blue line without any specific peak). C cell positive only for EF1α indicating neither a FSHβ (blue line) nor LHβ (black line) producing cell.

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