sequences encoding mruby2  (Addgene inc)

Journal: bioRxiv

Article Title: High and low exogenous nitrate concentrations produce distinct calcium signatures in Arabidopsis roots

doi: 10.1101/2025.03.03.641058

Figure Lengend Snippet: Asynchronous normalized Ca 2+ signal responses of the root hair (RH) compared to non-root hair (NRH) bearing epidermal cells showing distinct signatures comprising waves and spikes under low (0.25 mM) and high (5 mM) NO 3 - treatment. (A) The kinetics of apparent Ca 2+ signal represented through the GFP fluorescence intensity responding to low NO 3 - treatment. The root tissue imaged shows the apparent Ca 2+ signal through GFP fluorescence with mRuby2 RFP background and captures the response 90 seconds after the NO 3 - treatment. The peak normalized Ca 2+ signal intensity was captured around 190 seconds for all RH sections (RH-base, RH-mid and RH-tip) while NRH cell had peak normalized Ca 2+ signal intensity was around 240 seconds after low NO 3 - treatment. (B) The kinetics of apparent Ca 2+ signal responding to high NO 3 - treatment for the selected root sections. The root tissue imaged shows the apparent Ca 2+ signal through GFP fluorescence with mRuby2 RFP background and captures the response 110 seconds after the NO 3 - treatment. The peak normalized Ca 2+ signal intensity was captured around 120 seconds for RH-tip, and close to 140 seconds for RH-mid and RH-base sections.

Article Snippet: Sequences encoding mRuby2 (#40260, ( ) and GCaMP6s (#40753, ( ) were obtained from Addgene (Watertown, MA).

Techniques: Fluorescence

Journal: bioRxiv

Article Title: High and low exogenous nitrate concentrations produce distinct calcium signatures in Arabidopsis roots

doi: 10.1101/2025.03.03.641058

Figure Lengend Snippet: Asynchronous normalized Ca 2+ signal in the root sections in response to the low NO 3 - treatment. A representative root section for the low NO 3 - treated seedling is shown with the normalized Ca 2+ signal, where section S1 was distal to the root tip while S4 was proximal to the root tip. The root tissue imaged above shows the apparent Ca 2+ signal through GFP fluorescence with mRuby2 RFP background and captures the response 120 seconds after low NO 3 - treatment. The peak normalized Ca 2+ signal intensity was captured around 90 seconds for sections S1 and S2, while the peak intensity of the normalized Ca 2+ signal was close to 125 seconds for S3 and 60 seconds for S4.

Article Snippet: Sequences encoding mRuby2 (#40260, ( ) and GCaMP6s (#40753, ( ) were obtained from Addgene (Watertown, MA).

Techniques: Fluorescence

Journal: bioRxiv

Article Title: TOR signaling regulates GPCR levels on the plasma membrane and suppresses the Saccharomyces cerevisiae mating pathway

doi: 10.1101/2024.05.09.593412

Figure Lengend Snippet: ( A-D ) Representative images of cells expressing ( A ) Ste2-mEnvy, ( B ) Ste3-mEnvy, ( C ) Gpr1-mEnvy, and ( D ) Pma1-mRuby2 treated with or without 0.2µM rapamycin for 2h. Scale bars represent 4µm. ( E ) Quantification of Ste2-mEnvy (n > 200 cells), Ste3-mEnvy (n > 280 cells), Gpr1-mEnvy (n > 340 cells), and Pma1-mRuby2 (n > 250 cells) mean fluorescent intensities at the plasma membrane. Measurements were normalized to the mean of the untreated groups to produce percentages. Error bars represent ±SEM. * = p < 0.05, ** = p < 0.01, *** = p < 0.001. ( F ) β-galactosidase mating transcription assays of cell cultures treated with or without 0.2µM rapamycin for 2h (n > 10 colonies). Error bars represent ±SEM. ( G ) Table of EC 50 and E max responses to α-factor.

Article Snippet: The sequence for PMA1 was amplified from BY4741 genomic DNA, and the mRUBY2 sequence was amplified from pFA6a-link-yomRuby2-Kan (Addgene Plasmid #44953) ( ).

Techniques: Expressing, Clinical Proteomics, Membrane