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. 2019 Oct;60(13):4336–4345. doi: 10.1167/iovs.19-27865

Figure 1.

Figure 1

TSH and TSHR signaling stimulate proliferation in orbital fibroblasts. (A) OFs explanted from patients with TED (GOFs, black bars) or without TED (NOFs, clear bars) were treated with 1, 10, or 50 mU/mL TSH (or without TSH, untreated) for 48 hours. The nucleotide analog BrdU was added for 24 hours to measure DNA synthesis. After culture, cells were fixed and the BrdU label was detected by ELISA as described in the Methods section. TSH treatment resulted in a dose-dependent increase in OF proliferation. GOFs accumulated significantly more BrdU than NOFs at all TSH doses. Results are presented as means ± SEM from triplicate wells repeated in n = 4 NOF strains and n = 5 GOF strains. #P < 0.05, ##P< 0.01, ###P < 0.001 versus untreated samples. **P< 0.01, ***P < 0.001 in GOF versus NOF samples of the same treatment. (B) Expression of TSHR mRNA was detected by qPCR in untreated NOFs (n = 4) and GOFs (n = 5). TSHR mRNA levels were normalized to GAPDH mRNA and 18S rRNA levels. GOFs had ∼1.25-fold more TSHR mRNA than NOFs, *P < 0.05. Relevant information on patient samples is given in the Table.