pgfp b rs backbone vector  (OriGene)

Journal: iScience

Article Title: Nuclear respiratory factor-1 negatively regulates TGF-β1 and attenuates pulmonary fibrosis

doi: 10.1016/j.isci.2021.103535

Figure Lengend Snippet: ReCORM increases HO-1 and NRF-1 proteins and decreases pro-fibrotic proteins (A–D) C57BL/6 mice were injected with 3 consecutive doses of 1 mg/kg ReCORM or iCORM. Seven days post-BLM with or without ReCORM, lungs were excised and analyzed for HO-1, NRF-1, Nrf2, and PGC-1α protein expression measured by immunoblotting, with tubulin protein as internal standard (A-D). (E) Immunoblots show lung TGFβ1 protein levels on day 7 post-BLM, which are reduced by ReCORM. (F) Scatterplots of TGFβ1 densitometry levels. (G) Immunoblot of TGFβR1 protein levels on day 7 post BLM; ReCORM reduces the receptor levels in the lung. (H) Scatterplots of TGFβR1 densitometry levels. Experiments done in triplicate (n = 6/group); representative data shown). Semi-quantitative group data are expressed as mean ± SD.∗P < 0.05 compared ReCORM with the iCORM group. †P < 0.05 ReCORM + BLM vs BLM group).

Article Snippet: Cells were transfected in 6-well plates at 70%–80% confluence with pGFPCRS vector containing HO-1 oligonucleotide sequence for optimal suppression of HO-1 (Origene, TR30018), NRF-1 (Origene, TF517273) or CCN5 (Origene, TF511452).

Techniques: Injection, Expressing, Western Blot

Journal: iScience

Article Title: Nuclear respiratory factor-1 negatively regulates TGF-β1 and attenuates pulmonary fibrosis

doi: 10.1016/j.isci.2021.103535

Figure Lengend Snippet: Human lung fibroblasts (MRC5) and BLM induction of fibrotic markers (A and B) MRC5 cells transduced with sh-HO-1 or sh-NRF1 to silence these genes were then exposed to BLM for 18 h. Transcripts for CCN5 or SMAD7 were determined as seen in scatterplots. (C and D) Immunoblot of cells transfected with NS-ShRNA or ShRNA targeting NRF-1 (Sh-NRF-1). (E and F) ReCORM inhibits TGFβ1 signaling dependent on HO1 and NRF-1. MRC5 cells transduced with sh-HO-1 or sh-NRF1 to silence these genes were then exposed to TGFβ1for 18 h. Transcripts for HO-1 or NRF1 were determined as seen in scatterplots. (G) Responsiveness of overexpression or silencing of CCN5 with and without ReCORM to TGFβ1 induction of α-SMA mRNA levels in MRC5 cells. (H) Responsiveness to overexpression or silencing of NRF-1 with and without ReCORM to TGF-β induction of α-SMA mRNA levels in MRC5 cells. All experiments in quadruplicate representative data are shown. Quantitative data expressed as mean ± SD. ∗P < 0.05 ReCORM vs iCORM or †P < 0.05 ReCORM + BLM or ReCORM + TGFβ1 vs BLM or TGFβ1 groups.

Article Snippet: Cells were transfected in 6-well plates at 70%–80% confluence with pGFPCRS vector containing HO-1 oligonucleotide sequence for optimal suppression of HO-1 (Origene, TR30018), NRF-1 (Origene, TF517273) or CCN5 (Origene, TF511452).

Techniques: Transduction, Western Blot, Transfection, shRNA, Over Expression

Journal: iScience

Article Title: Nuclear respiratory factor-1 negatively regulates TGF-β1 and attenuates pulmonary fibrosis

doi: 10.1016/j.isci.2021.103535

Figure Lengend Snippet:

Article Snippet: Cells were transfected in 6-well plates at 70%–80% confluence with pGFPCRS vector containing HO-1 oligonucleotide sequence for optimal suppression of HO-1 (Origene, TR30018), NRF-1 (Origene, TF517273) or CCN5 (Origene, TF511452).

Techniques: Recombinant, Software, TUNEL Assay