Data Supplement

• Data Supplement -

  Supplemental Figure 1 - Influence of LY341495 on mGlu5 activity and responses to CDPPB in HEK293A cells.

  Supplemental Figure 2 - DHPG-induced real-time internalization of mGlu₅.

  Supplemental Figure 3 - Concentration-response relationships for mGlu₅ internalization kinetics induced by PAMs in the presence of orthosteric agonists (DHPG or L-glutamate) or antagonists (LY341495).

  Supplemental Figure 4 - Comparison of mGlu₅ PAM potencies and maximal responses in HEK293A cells across the four functional assays.

  Supplemental Figure 5 - Assessment of probe dependence of mGlu₅ PAMs. (A, B and D) No probe dependence was evident when comparing the E_max, pEC₅₀ and pK_B values obtained from PAM potentiation of L-glutamate and DHPG.

  Supplemental Figure 6 - Comparison of pK_I estimates from radioligand inhibition binding and pK_B values from functional assays for mGlu₅ PAMs.

  Supplemental Table 1 -  pEC₅₀ values from functional assays for orthosteric ligands in HEK293A cells or cortical neurons.

  Supplemental Table 2 - : CDPPB pEC₅₀ and E_max values from functional assays in HEK293A cells in the presence of 300 µM LY341495.

  Supplemental Table 3 - Potency and maximal potentiation estimates for mGlu₅ PAM potentiation of the indicated orthosteric agonist at ~EC₂₀ in HEK293A cells.

  supplemental Table 4 -  Potency and maximal response of mGlu₅ PAMs as agonists or potentiators of DHPG in cortical neurons.