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

Journal: Frontiers in Immunology

Article Title: Early sex-related transcriptional differences in CD8 + T cells responding to chronic viral infection reveal a sex bias in exhaustion

doi: 10.3389/fimmu.2026.1783098

Figure Lengend Snippet: Sex-related CD8 + T cell responses to LCMV-Armstrong and LCMV-clone 13 infection. (a) Experimental approach (Image created with biorender.com ). Flow cytometry staining of CD62L hi CD44 lo naïve T cells (gated on CD45.1 + CD8 + T cells) is shown. F, female; M, male; Arm, LCMV-Armstrong virus; Cl13, LCMV-clone 13 virus; FACS, fluorescence activated cell sorting; scRNA-seq, single-cell RNA-sequencing. (b) Absolute cell numbers of transferred CD45.1 + CD8 + T cells that are female (F, circles) or male (M, squares), analyzed in the spleen of LCMV-Armstrong (top) or LCMV-clone 13 (bottom) infected mice at 7 dpi. Summary of three independent experiments for LCMV-Armstrong ( n = 11 mice) and four independent experiments for LCMV-clone 13 ( n = 12 female mice, n = 14 male mice). Independent experiments are represented by different shades of orange (female) and blue (male). * p<0.05, significant, ns, not significant, determined using an unpaired two-tailed Student’s t -test. Arm, LCMV-Armstrong virus; Cl13, LCMV-clone 13 virus. (c) Absolute cell numbers of transferred CD45.1 + CD8 + T cells that are female (F, circles) or male (M, squares) in the blood at 7 dpi with LCMV-Armstrong (top) and LCMV-clone 13 (bottom) viruses. Independent experiments are represented by different shades of orange (female) and blue (male). Summary of three LCMV-Armstrong and four LCMV-clone 13 independent experiments with at least three mice per group. *** p<0.001, ns, not significant, determined using an unpaired two-tailed Student’s t -test. Arm, LCMV-Armstrong virus; Cl13, LCMV-clone 13 virus; dpi, days post-infection. (d) MSD, Meso Scale discovery analyses of pro-inflammatory cytokines in the sera of LCMV-Armstrong (top, closed circles, closed squares) or LCMV-clone 13 (bottom, open circles, open squares) infected F, female (circles) or M, male (squares) mice at 7 dpi. Summary of two independent experiments with five to twelve mice per group. Box and whiskers plots show minimum and maximum values and line at the median. Each symbol represents an individual mouse. Error bars indicate standard error of the mean (SEM). ** p<0.01, * p<0.05, ns, not significant, statistical significance of differences was analyzed between female and male samples per infection type, determined using unpaired two-tailed Student’s t -test.

Article Snippet: Barcoded single-cell (sc) cDNA libraries were generated (10X Genomics) from inactivated naïve and antigen-specific CD8 + T cells harvested at 7 dpi with LCMV-Armstrong or LCMV-clone 13 viruses.

Techniques: Infection, Flow Cytometry, Staining, Virus, Fluorescence, FACS, Single Cell, RNA Sequencing, Two Tailed Test