Figure 4
Figure 4. Expression dynamics of the most significantly regulated transcripts. (A) Key to the 5-way Venn diagram in Figure 4B of the 327 most significantly differentially expressed genes (DE_Sig) showing the overlap of fold change combinations. The 3 circles represent transcripts differentially expressed between Late-Es (L) and one of CFU-E (C, green), Pro-E (P, blue), or Int-E (I, red). The wide magenta arc represents genes differentially regulated between Pro-Es and Int-Es. The narrower yellow arc encompassing most of the perimeter of the wider arc represents genes differentially regulated between CFU-E and Int-Es. The colors merge where the shapes intersect each other. (B) Five-way Venn diagram of the 327 most significantly differentially expressed transcripts showing the overlap of fold change combinations (see panel A for key). The number of transcripts that are up-regulated during maturation are shown in white numbers with a dark shadow, whereas the number of transcripts that are down-regulated during maturation are shown in black numbers with a white shadow. Most genes are either terminally up-regulated or terminally down-regulated. For example, 98 transcripts are up-regulated between CFU-E to Late-Es, Pro-Es to Late-Es to Int-Es to Late-Es, and 92 transcripts are down-regulated between CFU-E to Late-Es, Pro-Es to Late-Es to Int-Es to Late-Es. One transcript is up-regulated between CFU-E to Int-Es and Pro-Es to Int-Es, but not between any of these stages and Late-Es. Where no number is shown, there were no transcripts in that set. (C) Heat maps of expression levels of transcripts most significantly differentially expressed (DE_Sig) up-regulated during erythroid maturation. Each column represents a different stage of erythroblast maturation, and each row represents a different PID. Red, white, and blue left-hand panels indicate log (base 2) of expression ratios between the stage indicated in the column heading and CFU-E during erythroid maturation. White represents a 1:1 ratio, so the CFU-E column is always white; red, up-regulation (log fold change of 3 indicating an 8-fold increase during maturation); and blue, down-regulation (log fold change of −3 indicating an 8-fold decrease during maturation). Red and blue right-hand panels indicate linear expression levels. Blue indicates no or low expression; and red, high expression (> 1000). Expression dynamics are indicated. (D) Heat maps of expression levels of transcripts most significantly differentially expressed (DE_Sig) down-regulated during erythroid maturation. Each column represents a different stage of erythroblast maturation, and each row represents a different PID. The red, white, and blue left-hand panel indicates log (base 2) of expression ratios between the stage indicated in the column heading and CFU-E during erythroid maturation. White represents a 1:1 ratio, so the CFU-E column is always white; red indicates up-regulation (log fold change of 3 indicating an 8-fold increase during maturation); and blue indicates down-regulation (log fold change of −3 indicating an 8-fold decrease during maturation). The red and blue right-hand panel indicates linear expression levels. Blue indicates no or low expression; and red, high expression (> 1000). Expression dynamics are indicated. (E) Significant increase in the range and magnitude of fold changes of expression of the 327 most differentially expressed transcripts (DE_Sig) from CFU-E/Pro-E (C-P), CFU-E/Int-E (C-I), CFU-E/Late-E (C-L), Pro-E/Int-E (P-I), Pro-E/Late-E (P-L), or Int-E/Late-E (I-L) in RNA from sorted erythroblasts (S) compared with that from unsorted cells (U). A 2-tailed paired t test was performed. Between each pair of stages compared, the detected fold changes were distributed over a narrower range of values in the unsorted erythroblasts compared with their sorted counterparts, indicating a dampening of the variation detected between different stages of erythroblasts in the unsorted samples.

Expression dynamics of the most significantly regulated transcripts. (A) Key to the 5-way Venn diagram in Figure 4B of the 327 most significantly differentially expressed genes (DE_Sig) showing the overlap of fold change combinations. The 3 circles represent transcripts differentially expressed between Late-Es (L) and one of CFU-E (C, green), Pro-E (P, blue), or Int-E (I, red). The wide magenta arc represents genes differentially regulated between Pro-Es and Int-Es. The narrower yellow arc encompassing most of the perimeter of the wider arc represents genes differentially regulated between CFU-E and Int-Es. The colors merge where the shapes intersect each other. (B) Five-way Venn diagram of the 327 most significantly differentially expressed transcripts showing the overlap of fold change combinations (see panel A for key). The number of transcripts that are up-regulated during maturation are shown in white numbers with a dark shadow, whereas the number of transcripts that are down-regulated during maturation are shown in black numbers with a white shadow. Most genes are either terminally up-regulated or terminally down-regulated. For example, 98 transcripts are up-regulated between CFU-E to Late-Es, Pro-Es to Late-Es to Int-Es to Late-Es, and 92 transcripts are down-regulated between CFU-E to Late-Es, Pro-Es to Late-Es to Int-Es to Late-Es. One transcript is up-regulated between CFU-E to Int-Es and Pro-Es to Int-Es, but not between any of these stages and Late-Es. Where no number is shown, there were no transcripts in that set. (C) Heat maps of expression levels of transcripts most significantly differentially expressed (DE_Sig) up-regulated during erythroid maturation. Each column represents a different stage of erythroblast maturation, and each row represents a different PID. Red, white, and blue left-hand panels indicate log (base 2) of expression ratios between the stage indicated in the column heading and CFU-E during erythroid maturation. White represents a 1:1 ratio, so the CFU-E column is always white; red, up-regulation (log fold change of 3 indicating an 8-fold increase during maturation); and blue, down-regulation (log fold change of −3 indicating an 8-fold decrease during maturation). Red and blue right-hand panels indicate linear expression levels. Blue indicates no or low expression; and red, high expression (> 1000). Expression dynamics are indicated. (D) Heat maps of expression levels of transcripts most significantly differentially expressed (DE_Sig) down-regulated during erythroid maturation. Each column represents a different stage of erythroblast maturation, and each row represents a different PID. The red, white, and blue left-hand panel indicates log (base 2) of expression ratios between the stage indicated in the column heading and CFU-E during erythroid maturation. White represents a 1:1 ratio, so the CFU-E column is always white; red indicates up-regulation (log fold change of 3 indicating an 8-fold increase during maturation); and blue indicates down-regulation (log fold change of −3 indicating an 8-fold decrease during maturation). The red and blue right-hand panel indicates linear expression levels. Blue indicates no or low expression; and red, high expression (> 1000). Expression dynamics are indicated. (E) Significant increase in the range and magnitude of fold changes of expression of the 327 most differentially expressed transcripts (DE_Sig) from CFU-E/Pro-E (C-P), CFU-E/Int-E (C-I), CFU-E/Late-E (C-L), Pro-E/Int-E (P-I), Pro-E/Late-E (P-L), or Int-E/Late-E (I-L) in RNA from sorted erythroblasts (S) compared with that from unsorted cells (U). A 2-tailed paired t test was performed. Between each pair of stages compared, the detected fold changes were distributed over a narrower range of values in the unsorted erythroblasts compared with their sorted counterparts, indicating a dampening of the variation detected between different stages of erythroblasts in the unsorted samples.

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