Skip to main content
. 2022 Feb 18;12:2847. doi: 10.1038/s41598-022-06678-7

Figure 2.

Figure 2

PAD2 regulates myofibroblast differentiation and extracellular matrix production. (A) Control (n = 4) and RA-ILD (n = 4) fibroblasts were treated with vehicle or bb-chloride-amidine (1 μM). 24 h later, cells were lysed and subjected to Western blot analysis to measure α-SMA expression. (B) Cells were mixed with collagen 1 solution as described in the Methods. Gel size was measured at 0 and 24 h after collagen gelation (n = 4 for each condition). (C) Cells were treated as described in (A). After treatment, cells were lysed for total RNA isolation. mRNA levels of collagen1a1 (COL1A1) and fibronectin (FN) were measured by RT-PCR. (D-E) Control (n = 4) and RA-ILD (n = 4) fibroblasts were transfected with scrambled (scr) or shPAD2 via lentivirus as described in the Methods. Stably transfected cells were subjected to Western blot analysis to measure (D) α-SMA and PAD2 expression, (E) cell contraction, and (F) RT-PCR to measure COL1A1 and FN. Data are mean ± SEM. P < 0.05; significant comparisons by one-way ANOVA: *vs. control, †vs. RA-ILD fibroblasts treated with vehicle or Scr. Abbreviations: α-SMA = alpha smooth muscle actin, GAPDH = glyceraldehyde 3-phosphate dehydrogenase, ILD = interstitial lung disease, PAD2 = peptidylarginine deminase 2, RA = rheumatoid arthritis, RT-PCR = real-time polymerase chain reaction, scr = scrambled short hairpin RNA, shPAD2 = PAD2 short hairpin RNA.