Fig 5. CSP-6 associates with WHI-2 in vivo, and loss of whi-2 results in phenotypes similar to Δcsp-6.

A: Silver stained SDS-PAGE gel showing CSP-6 associated with WHI-2 (band 5) as confirmed by MS-based protein sequencing. Bands 1–4 correspond to CSP-6 itself based on Mass Spec data (S2 Table). B: Co-immunoprecipitation assay showing CSP-6 physically interacts with WHI-2 in vivo. A wild type strain (ras-1^bd) that lacks the epitope tag was used as a negative control. Two individual transformants (-10 and -13) of csp-6^FLAG, whi-2^V5 were used in Co-IPs and Western analyses. C: Cultures of WT (ras-1^bd or 74A), Δcsp-6 and Δwhi-2 (with or without ras-1^bd respectively) were grown on solid minimal slants showing similar growth morphology between Δcsp-6 and Δwhi-2. D: Race tube data showing loss of overt conidiation rhythms in Δwhi-2; triplicates of each strain are shown. E: Luciferase traces showing frq-luc was circadianly rhythmic but expressed with a delayed phase in the Δwhi-2 mutant. ‘p’ stands for period. F: Luciferase activity assay of frq C-box-luc in strains of ras-1^bd, the Δcsp-6, ras-1^bd strain, the Δwhi-2, ras-1^bd strain, and double mutant Δcsp-6, Δwhi-2, ras-1^bd strain, showing the phase delay seen in Δcsp-6 and in the double mutant Δcsp-6, Δwhi-2 was similar.