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. 2020 Feb 13;5(3):e133972. doi: 10.1172/jci.insight.133972

Figure 4. Stiff matrix promotes APA of COL1A1 and FN1 in favor of the proximal poly(A) site usage.

Figure 4

(A) A schematic diagram of the COL1A1 or FN1 transcripts, in which the relative positions of complete coding sequence (CDS), proximal and distal poly(A) sites, and primer pairs for amplification of a common region and a distal region in the long transcripts are shown. (B–G) Lung fibroblasts were cultured on soft and stiff hydrogels for 48 hours. Relative levels of total/common COL1A1 transcripts (B), long/distal COL1A1 transcripts (C), total/common FN1 transcripts (E), and long/distal FN1 transcripts (F) were determined by real-time RT-PCR. GAPDH was used as internal reference control. Ratios of long-to-total COL1A1 (D) and FN1 (G) transcripts were used to indirectly evaluate the proximal poly(A) site usage. Bar graphs represent (mean ± SD) 5 separate experiments. A 2-tailed Student’s t test was used for comparison between groups.