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. 2020 Oct 13;24:100825. doi: 10.1016/j.bbrep.2020.100825

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© 2020 The Author(s)

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Fig. 4 — Effect of 1,25D and UroA and/or Res on TPH2 mRNA concentrations in serotonergic rat raphe RN46A-B14 cells as well as on total serotonin (5-HT) levels in the neuronal cell culture medium. (A) Expression of TPH2 in serotonergic rat raphe RN46A-B14 cells treated with 1,25D (10 nM) and UroA (10 μM) and/or Res (25 μM). Each mRNA value is the average of 19 biological replicates ± Std. Dev. Fold-effect over EtOH/vehicle control is listed at the top of each bar. Only UroA and UroA + Res are capable of statistically significantly elevating TPH2 mRNA level above that achieved by 1,25D treatment alone. (B) Serotonin (5-HT) levels in low basal serotonin (0.55 ng/ml) RN46A-B14 cell culture medium in response to treatment with 1,25D (10 nM) and UroA (10 μM) and/or Res (25 μM). A 200 μL aliquot of medium was removed from each plate after 72 h of treatment and 5-HT quantitated by ELISA as detailed in Methods. Values are the average of three biological replicates (n = 3) ± Std. Dev., with duplicate or triplicate assay samples averaged in each group. Fold-effect over EtOH/vehicle control is listed at the top of each bar. Employing ANOVA, only UroA is capable of statistically significantly (p = 0.0015) elevating 5-HT level above that achieved by 1,25D treatment alone; all other values are not statistically significantly different from their respective controls. (C) Serotonin (5-HT) levels in high basal serotonin (1.23 ng/ml) RN46A-B14 cell culture medium in response to treatment with 1,25D (10 nM) and UroA (10 μM) and/or Res (25 μM). A 200 μL aliquot of medium was removed from each plate after 72 h of treatment and 5-HT quantitated by ELISA as detailed in Methods. Values are the average of three biological replicates (n = 3) ± Std. Dev., with duplicate or triplicate assay samples averaged in each group. Fold-effect over EtOH/vehicle control is listed at the top of each bar. Employing ANOVA, only UroA is capable of statistically significantly (p = 0.0003) elevating 5-HT level above that achieved by 1,25D treatment alone; all other values are not statistically significantly different from their respective controls.

Fig. 4 — Effect of 1,25D and UroA and/or Res on TPH2 mRNA concentrations in serotonergic rat raphe RN46A-B14 cells as well as on total serotonin (5-HT) levels in the neuronal cell culture medium. (A) Expression of TPH2 in serotonergic rat raphe RN46A-B14 cells treated with 1,25D (10 nM) and UroA (10 μM) and/or Res (25 μM). Each mRNA value is the average of 19 biological replicates ± Std. Dev. Fold-effect over EtOH/vehicle control is listed at the top of each bar. Only UroA and UroA + Res are capable of statistically significantly elevating TPH2 mRNA level above that achieved by 1,25D treatment alone. (B) Serotonin (5-HT) levels in low basal serotonin (0.55 ng/ml) RN46A-B14 cell culture medium in response to treatment with 1,25D (10 nM) and UroA (10 μM) and/or Res (25 μM). A 200 μL aliquot of medium was removed from each plate after 72 h of treatment and 5-HT quantitated by ELISA as detailed in Methods. Values are the average of three biological replicates (n = 3) ± Std. Dev., with duplicate or triplicate assay samples averaged in each group. Fold-effect over EtOH/vehicle control is listed at the top of each bar. Employing ANOVA, only UroA is capable of statistically significantly (p = 0.0015) elevating 5-HT level above that achieved by 1,25D treatment alone; all other values are not statistically significantly different from their respective controls. (C) Serotonin (5-HT) levels in high basal serotonin (1.23 ng/ml) RN46A-B14 cell culture medium in response to treatment with 1,25D (10 nM) and UroA (10 μM) and/or Res (25 μM). A 200 μL aliquot of medium was removed from each plate after 72 h of treatment and 5-HT quantitated by ELISA as detailed in Methods. Values are the average of three biological replicates (n = 3) ± Std. Dev., with duplicate or triplicate assay samples averaged in each group. Fold-effect over EtOH/vehicle control is listed at the top of each bar. Employing ANOVA, only UroA is capable of statistically significantly (p = 0.0003) elevating 5-HT level above that achieved by 1,25D treatment alone; all other values are not statistically significantly different from their respective controls.