Fig. 6. Processing of TLC1 3′ ends is not altered in pop cells at 24 °C.

The 3′ end sequences of TLC1 extracted from 24 or 30 °C grown cells were analyzed by 3′ RACE. a Diagram of the 3′ end of TLC1 from nucleotides 1139 to 1164. The last nucleotide in the 1157 nt mature TLC1 molecule is indicated with a red arrowhead. b PCR products amplified from TLC1 cDNA were analyzed on a 1% agarose gel. The expected sizes of mature and precursor forms of TLC1, are, respectively, ~220 and ~350 bp. Compared to WT, the relatively abundance of short to long forms were higher in pop1 and pop6 cells at 24 °C and, especially, 30 °C. Ratios of long to mature form are shown below lanes. All amplicons <500 nts shown in panel B were purified and deep sequenced. The relative abundance of each TLC1 species as determined by sequencing was quantified. Sequences that aligned to the 1154 to >1159 nt end of TLC1 with or without a poly(A) tail are shown for cells grown at c 24 °C or d 30 °C. The size of the poly(A) tails are: no adenosine (gray), 1 adenosine (orange), 2-5 adenosines (dark blue), 6–10 adenosines (yellow) and >10 adenosines (green). Total reads are indicated. Each bar shows the average from biological duplicates. At 30 °C, transcripts ending at nt 1156 [no poly(A)] were significantly more abundant by an unpaired two-tailed Student’s t-test in pop1 and pop6 versus WT (marked with asterisk). Source data are provided as a Source Data file.