(Solved): Figure 1. Immunoblot analyses of different proteins. In the immunoblots represented, lanes \( ...

Figure 1. Immunoblot analyses of different proteins. In the immunoblots represented, lanes \( 1,10,17,22 \), and 26 represent molecular weight markers. Samples \( 2-9 \) were immunoblotted with an antibody against Protein 1; samples 11-16 were immunoblotted against Protein 2; samples 18-21 were immunoblotted against Protein 3; and samples 23-26 were immunoblotted against Protein 4. For Protein 1, the cells represented in lanes 2 and 6 were grown in normal culture media, the cells in lanes 3-5 were grown in the presence of increasing concentrations of Chemical A, and the cells in lanes \( 7-9 \) were grown in the presence of increasing concentrations of Chemical B. In all cases the cells were collected after 48 hours of exposure to the growth conditions indicated. For Protein 2, two different versions of the protein were expressed by transfection in cells that do not normally express that protein. The WT form (wild-type) is the original (unaltered) primary sequence of the protein, whereas the \( \Delta N \) (deletion at the \( N \)-terminus) is a mutant form of the protein in which the first \( \mathrm{N} \)-terminal 25 amino acid residues were deleted. Lanes 11 and 12 represent the two forms of the protein expressed in cells grown under normal conditions, whereas lanes 13-16 represent the proteins expressed in cells exposed to heat-shock conditions \( \left(43^{\circ} \mathrm{C}\right. \) for 2 hours). In lanes 15 and 16, the cells were also treated with MG132 (a proteasomal inhibitor). Serine and Threonine phosphorylation sites were removed; ii) the \( \Delta Y \odot \), in which al predicted Tyrosine phosphorylation sites were removed; and, iii) the \( \triangle \mathrm{CT} \), in which the last 4 amino acid residues at the \( \mathrm{C}- \) terminus were removed, including a Cysteine residue located right before the last 3 amino acid residues. All of these mutant forms, as well as the original wild-type (WT) form of the protein were expressed by transfection into a cell line that normally doesn't express this protein. For Protein 4, two different mutant forms of the protein were developed: K41A, in which a Lysine residue located in position 41 was substituted with Alanine, and K90A, in which a Lysine residue located in position 90 was substituted with Alanine. These two mutants were developed because those Lysine residues were located within consensus sequences predicted to be modified by a type of post-translational modification that could explain the observed upper band for that protein (the predicted molecular weight for that protein is 35 kDa). Those mutant forms, as well as the original wild-type (WT) form of the protein were expressed by transfection into a cell line that normally doesn't express this protein. "(This question is related to Protein 4) In Fig. 1, the overall profile observed for Protein 4 indicates that this protein is modified by " "(This question is related to Protein 2) In Fig. 1, the overall profile observed for Protein 2 indicates that when the cells are expossed to heat-shock conditions, Protein 2" is retained in the nucleus is retained in the cytoplasm is secreted out of the cell the protein continues to be targeted to its original cellular compartment that it is targeted to in its wild-type form becomes rapidly degraded by a ubiquitin-dependent mechanism is heavily phosphorylated "(This question is related to Protein 1) In Fig. 1, if Chemical B is known to block the activity of a Ubiquitin ligase, it would be fair to state that" the type of Ubiquitin ligase affected by Chemical B has the ability to poly-ubiquitinylate Protein 1 the type of Ubiquitin ligase affected by Chemical B does not recognize Protein 1 Protein 1 is not poly-ubiquitinylated Chemical B triggers the de-Ubiquitinylation of Protein 1 "(This question is related to Protein 2) In Fig. 1, the overall profile observed for Protein 2 indicates that when the N-terminal 25 amino acid residues are eliminated, Protein 2" is retained in the nucleus is retained in the cytoplasm is secreted out of the cell the protein continues to be targeted to its original cellular compartment that it is targeted to in its wild-type form becomes rapidly degraded by a ubiquitin-dependent mechanism aggregates