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

Journal: PLoS ONE

Article Title: Integration of Transcriptomics, Proteomics, and MicroRNA Analyses Reveals Novel MicroRNA Regulation of Targets in the Mammalian Inner Ear

doi: 10.1371/journal.pone.0018195

Figure Lengend Snippet: ( A ) Schematic illustrations of a P0 whole inner ear, cochlea and vestibule: utricle (ut), saccule (sa), spiral ganglia (sg), scala media (sm), scala vestibule (sv), scala tympani (st). In purple, cochlear hair cells (chc) and vestibular hair cells (vhc). ( B-D ) Specific expression patterns were demonstrated for the miRNAs by ISH in whole mount inner ears, followed by cryosectioning. For each miRNA images of the whole inner ear (top) and magnified cochlea (middle) and vestibule (utricle or saccule, bottom) are shown separately. ( B ) A probe for miR-182 was used as a positive control and confirmed probe penetration and staining throughout the inner ear. miR-182 is expressed in the inner ear hair cells and spiral ganglia at this age . Expression patterns for miR-135b ( C ) and miR-205 ( D ) were consistent with the miRNA array analysis. Scale bars: 100 µm.

Article Snippet: Total RNA was converted to single stranded cDNA using the High Capacity cDNA Reverse Transcription Kit (Applied Biosystems). qRT-PCR was performed using Taqman Gene Expression Assay for Psip1 and either Tbp or RPLP0 and the 2X Universal PCR Master Mix (Applied Biosystems).

Techniques: Expressing, Positive Control, Staining