Fig 2: TXNDC5 was upregulated in IPF lungs/lung fibroblasts and correlated with fibrogenic genes.a Transcript analysis showed significant upregulation of TXNDC5 mRNA in human IPF (n = 26, central and lateral lung from 13 biologically independent samples), compared with control (n = 18, central and lateral lung from 9 biologically independent samples), lung tissues. b Immunoblot analysis showed a marked increase in TXNDC5 and aSMA protein levels in human IPF (n = 5 biologically independent samples), compared with control (n = 3 biologically independent samples), lung tissues. c Protein expression level of TXNDC5 and aSMA were also increased in IPF, compared with control, lung fibroblasts following TGFß1 treatment (n = 5 biologically independent samples per group). d Re-analyses of microarray data from IPF human lung fibroblasts (GSE40839)15 revealed strong positive correlation between the expression level of TXNDC5 and that of genes encoding fibrogenic proteins (COL1A1, ELN, ACTA2) in human lung fibroblasts (n = 13 biologically independent samples per group) (Data are presented as mean ± SEM, P value determined using two-tailed Mann–Whitney U test. Source data are provided as a Source Data file).

Fig 3: Maternal transmission of antimiR-29a-3p to mouse pups increases steady-state expression levels of miR-29a-3p target-genes. Either a scrambled antimiR (open symbols) or an antimiR directed against miR-29a-3p (closed symbols) was administered by intravenous (i.v.) injection at a dose of 100 mg kg-1 to pregnant dams on the 17th day of gestation. The liver was harvested from mouse pups on postnatal day (P)7. Steady-state mRNA expression levels were assessed by real-time RT-PCR for the miR-29a-3p targets (A) Eln, and (C) Adamts7, described by the mean ?CT ± SD. (n = 4–6 animals per group; each symbol represents an animal, where some symbols may coalesce), where ?CT = CT(Polr2a) - CT(gene of interest). Mean values were compared between treatment conditions by unpaired Student's t-test. P-values are provided when P < 0.05. Total liver protein extracts from pups at P7 were screened by immunoblot for steady-state protein expression levels of (B) elastin (ELN) and (D) ADAM metallopeptidase with thrombospondin type 1 motif 7 (ADAMTS7), where ß-ACTIN served as a loading control. For immunoblots, protein extracts from three scrambled antimiR-treated mice and three antimiR-29a-3p-treated mice were run side-by-side, on the same gel.

Fig 4: TXNDC5 was upregulated in BLM-induced PF and enriched in lung fibroblasts in vivo.a Transcript expression levels of Txndc5 and fibrogenic protein genes (Col1a1, Col3a1, Eln, Fn and Ctgf) were markedly increased in the lung tissues from BLM-(n = 4 biologically independent animals per group), compared with PBS- (Sham, n = 3 biologically independent animals), treated WT mice on Day 21. b Immunohistochemical (IHC) staining on the serial sections of mouse lungs showed a strong upregulation of aSMA and TXNDC5 in the lung tissues from BLM-, compared with PBS- (Sham), treated WT mice on Day 21 (n = 9 fields examined over 3 biologically independent animals per group). c IF staining for TXNDC5 on lung sections from sham-operated and BLM-treated Col1a1-GFPTg mice. BLM treatment significantly increased TXNDC5 expression and the number of GFP-positive cells in the mouse lungs on Day 21 (n = 3 biologically independent animals). There was a high degree of co-localization of TXNDC5 with GFP-positive, collagen producing lung fibroblasts (white arrows and inset) (data are presented as mean SEM, P value determined using two-tailed Mann–Whitney U test. Source data are provided as a Source Data file. BLM bleomycin).