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

Figure 3.

Figure 3

Syndecan-2 (SDC2) inhibits α-SMA and PAD2 expression by regulating CD148/PI3K/Akt signaling in RA-ILD fibroblasts. Control (n = 4) and RA-ILD (n = 4) fibroblasts were transfected with scramble (scr) or shCD148 via lentivirus as described in the Methods. Stably transfected cells were treated with syndecan-2 (10 μg/ml) for 24 h. After treatment, cells were subjected to (A) Western blot to measure phospho-Akt, α-SMA, and PAD2 expression, (B) RT-PCR to measure mRNA levels of PADI2 (PAD2), or (C) a Sp1 transcriptional activity assay as described in the Methods. (D) RA-ILD fibroblasts were transfected via lentivirus with empty vector or pLenti-GIII-CMV-C-term-HA plasmid carrying the PADI2 (PAD2-over). Stably transfected cells were treated with syndecan-2 (10 μM) for 24 h. After treatment, cells were subjected to RT-PCR to measure mRNA levels of COL1A1 and PADI2. Data are mean ± SEM. P < 0.05; significant comparisons by one-way ANOVA: * vs. control, † vs. RA-ILD fibroblasts + Scr, # vs. RA-ILD fibroblasts + Scr + SDC2. Abbreviations: α-SMA = alpha-smooth muscle actin, ILD = interstitial lung disease, p-Akt = phosphorylated Akt, PAD2 = peptidylarginine deiminase 2, PI3K = phosphoinositide 3-kinase, RA = rheumatoid arthritis, scr = scrambled short hairpin RNA, SDC2 = syndecan-2, shCD148 = CD148 short hairpin RNA, shSp1 = Sp1 short hairpin RNA.