Journal: bioRxiv

Article Title: Molecular cloning of a novel, nervous system-specific RGS6 isoform lacking canonical G protein regulatory effects and with dominant negative actions

doi: 10.64898/2026.05.08.723811

Figure Lengend Snippet: Molecular cloning of novel RGS6 splice forms from human brain. ( A ) Splicing diagram of known RGS6 splice forms. Location of primers used for PCR-based amplification of RGS6 transcripts from a human whole brain cDNA library are shown in red. ( B ) Representative plasmid restriction digest (right) or nested PCR (down) indicating positive hits whose size is larger than known RGS6 splice forms. Each gel has a confirmed RGS6Lα1(+GGL) transcript (largest known splice form) indicated as well as yellow asterisks denoting transcripts encoding RGS6LA3α1(+GGL) which encodes RGS6B.

Article Snippet: Full-length cDNAs encoding novel RGS6 splice forms were amplified from a Marathon ready human brain cDNA library (Clontech; Mountain View, CA, USA) using a PCR-based strategy.

Techniques: Molecular Cloning, Amplification, cDNA Library Assay, Plasmid Preparation, Nested PCR

Journal: bioRxiv

Article Title: Molecular cloning of a novel, nervous system-specific RGS6 isoform lacking canonical G protein regulatory effects and with dominant negative actions

doi: 10.64898/2026.05.08.723811

Figure Lengend Snippet: RGS6B is an RGS6L(+GGL) isoform containing A3 and the α terminal exon (RGS6LA3α1(+GGL)). ( A ) Schematic diagram of the exon splicing scheme comparing RGS6Lα1(+GGL) (RGS6L) and RGS6LA3α1(+GGL) (RGS6B). mRNA sequence conservation of exon A3 between mouse and human is depicted below with the consensus sequence from 100 vertebrate species extracted by PhyloP. PCR amplification of exon A3 containing mRNA transcripts from select mouse tissues ( B ) or human cDNA libraries ( C ). Plasmids encoding RGS6Lα1, RGS6Lα2 and RGS6LA3α1 (all +GGL) are used as negative and positive controls, respectively. LNG, lung; KDN, kidney; LVR, liver; SPL, spleen; CTX, cortex; MB, midbrain; CRB, cerebellum; HIP, hippocampus; MSC, muscle; HRT, heart; PNC, peripheral nervous system; BR, whole brain. ( D ) HEK293T cells were transfected with a plasmid encoding RGS6LA3α1(+GGL) to confirm co-migration with mouse RGS6B via immunoblotting. ( E ) shRNA or miRNA constructs targeting the A3 exon were introduced into mouse primary cortical astrocytes. The ratio of RGS6B: RGS6L was determined via immunoblot and quantified from 4 independent experiments. α Tubulin serves as a loading control for immunoblots. Data were analyzed by one sample t-test to detect deviation of each group from 100% (scramble RNAi control). *P<0.05, ***P<0.001. Data are expressed as mean ± S.E.M.

Article Snippet: Full-length cDNAs encoding novel RGS6 splice forms were amplified from a Marathon ready human brain cDNA library (Clontech; Mountain View, CA, USA) using a PCR-based strategy.

Techniques: Sequencing, Amplification, Transfection, Plasmid Preparation, Migration, Western Blot, shRNA, Construct, Control