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. 2019 Apr;25(4):496–506. doi: 10.1261/rna.067967.118

FIGURE 4.

FIGURE 4.

Effect of Bcd1p on the molecular associations of several snoRNP-related proteins. (A–D) Co-IP analyses were performed in RSA1-TAP X GAL1::3HA-BCD1 (A,C,D) and PIH1-TAP X GAL1::3HA-BCD1 (B) cells transformed with a p416 vector expressing a Flag-tagged version of Snu13p (A), Nop58p (B,D), and Pih1p (C), or an empty p416 vector as control. Yeast cells were maintained in YPG medium or shifted to YPD medium for 6 h and all IPs were performed with an anti-Flag antibody. When useful, a second IP was performed by adjusting the cell extract volume to the change in protein expression level induced by the shift in the medium (in B,D) in order to minimize this effect. The Dps1 protein was used to control protein loading. The relative interactions in presence (white drawbars) or absence (black drawbars) of Bcd1p were quantified in three independent experiments. (E) ChIP assays were performed on GAL1::3HA-BCD1, GAL1::3HA-BCD1 x rsa1Δ or GAL1::3HA-BCD1 x pih1Δ cells transformed with a p416 vector expressing a Flag-tagged version of Nop58p. The cells were maintained in YPG medium or shifted to YPD medium for 6 h, and IP was performed with anti-Flag beads or GSH beads as control.