Anti rarα antibody

We used pSilencer 3.1-H1 (Life Technologies) as an shRNA vector. The sequences of the shRNA duplexes used RARα knockdown were as follows: RARα gene, sense 5′- GCAAGTACACTACGAACAACATTCAAGAGATGTTGTTCGTAGTGTACTTGCTTTTTTGGAA-3′ and antisense 5′- TTCCAAAAAAGCAAGTACACTACGAACAACATCTCTTGAATGTTGTTCGTAGTGTACTTGC-3′, no-target as a control, sense 5′-GTACTATTCGACACGCGAAGTTCAAGAGACTTCGCGTGTCGAATAGTACTTTTTTGGAA-3′ and antisense 5′-TTCCAAAAAAGTACTATTCGACACGCGAAGTCTCTTGAACTTCGCGTGTCGAATAGTAC-3′. Lipofectamine 2000 was used to transfect with each of these constructs into P19 cells. Western blotting probed with an anti-RARα antibody (Santa Cruz Biotechnology, CA) and anti-Beta-Actin antibody (Thermo SCIENTIFIC, CA) were performed to assess RARα knockdown. The influence of Tal2 expression by RARα suppression was examined by real-time PCR.

Primary antibodies for Western blot analysis were as follows: anti-LOX1 antibody (Santa Cruz Biotechnology, Santa Cruz, CA), anti-STRA6 antibody (ABGENT, San Diego, CA), anti-CRBP1 antibody (Santa Cruz Biotechnology), anti-RARα antibody (Santa Cruz Biotechnology), anti-RARγ antibody (Santa Cruz Biotechnology), anti-RXRα antibody (Santa Cruz Biotechnology), anti-c-Jun N-terminal kinase (JNK) antibody (Santa Cruz Biotechnology), anti-pJNK antibody (Abcam, Cambridge, MA), anti-p38MAPK antibody (ABGENT), anti-p-p38MAPK antibody (ABGENT), anti-pSmad2 antibody (Santa Cruz Biotechnology), anti-Smad2 antibody (Santa Cruz Biotechnology), anti-TGFβ₁ (Santa Cruz Biotechnology), anti-caspase 3 antibody (Santa Cruz Biotechnology), anti-collagen 1 antibody (Santa Cruz Biotechnology), and anti-actin antibody (Millipore, Temecula, CA). Secondary antibodies for Western blot analysis as HRP-conjugate antibody were purchased from Millipore. The inhibitors of JNK, SP600125 and p38MAPK, SB203580 were purchased from Sigma-Aldrich (St. Louis, MO).