Journal: Scientific Reports
Article Title: TGF-β1 increases viral burden and promotes HIV-1 latency in primary differentiated human bronchial epithelial cells
doi: 10.1038/s41598-019-49056-6
Figure Lengend Snippet: TGF-β signaling alters mRNA expression of HIV-1 host restriction factors in NHBE cells. NHBE ALI cultures were treated with recombinant TGF-β1 (10 ng/ml; vehicle as control). 48 hours post-treatment, total RNA was isolated and mRNA levels of IFITM3, PSIP1 and BLIMP-1 were determined by qRT-PCR. ( a ) TGF-β1 does not alter expression of IFITM3 mRNA. ( b ) TGF-β1 suppresses PSIP1 mRNA. ( c ) TGF-β1 increases mRNA levels of BLIMP-1 compared to vehicle treated controls. n = NHBE ALI cultures from 3 different lungs; *significant (p < 0.05).
Article Snippet: To examine mRNA expression, total RNA was extracted using the Qiagen RNeasy mini kit (Cat # 74104) and complementary DNA (cDNA) was reverse transcribed using the Applied Biosystems high-capacity cDNA reverse transcription kit (Cat # 4368814). qRT-PCR was performed on the Bio-Rad CFX96 real-time system using validated TaqMan probes (Life Technologies/Applied Biosystems HIV-LTR, Cat # Pa03453409_s1; IFITM3, Cat # Hs03057129_s1; BST2, Cat # Hs01561315_m1; BLIMP-1, Cat # Hs00153357_m1; PSIP1, Cat # Hs01045711_g1; and GAPDH, Cat # Hs02786624_g1). qRT-PCR results are represented as relative quantification normalized against internal control (GAPDH).
Techniques: Expressing, Recombinant, Control, Isolation, Quantitative RT-PCR