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. 2006 Jul 18;4(8):e232. doi: 10.1371/journal.pbio.0040232

Figure 5. Differentially Modulated Ptch1 Action in Dhcr7 –/– and Sc5d –/– MEFs .

Figure 5

(A) Basal Gli reporter activity measured in MEFs either deficient for Sc5d or Dhcr7 shows that the stacking of 7-DHC (in the Dhcr7 –/– MEFs) lowers Gli activity in the mutant cells compared with lathosterol stacking in the Sc5d –/– MEFs. Transfer of the conditioned media to reporter cells (indicated as “MEFs as medium donors”) shows that the inhibitory potential of the Dhcr7 –/– MEFs is medium-borne. Media were incubated on donor MEFs for 16 h and transferred to reporter cells for 6 to 8 h. ( n ≥ 4; *, p < 0.05; ***, p < 0.005).

(B) Medium transfer of Ptch1, Ptch1 siRNA, or control-transfected Sc5d –/– and Dhcr7 –/– MEFs shows that cells stacking lathosterol but lacking 7-DHC ( Sc5d –/– MEFs) are not able to translate different Ptch1 levels to an inhibitory action on reporter cells. The Dhcr7 –/– cells were able to properly convey an inhibitory signal when transfected with Ptch1 or, inversely, show a diminished inhibitory potential upon Ptch1 siRNA transfection. UVB treatment of Ptch1 transfectant medium (2 h) amplified the inhibitory action. Media incubations and statistics are as in (A).