Journal: STAR Protocols

Article Title: Protocol for high-resolution 3D spatial transcriptomics using Open-ST

doi: 10.1016/j.xpro.2024.103521

Figure Lengend Snippet:

Article Snippet: Alkaline phosphatase calf intestinal (CIAP) enzyme , Promega , Cat#M1821.

Techniques: Recombinant, Sterility, Reverse Transcription, Marker, Electrophoresis, Purification, Sequencing, Software, Microarray, Real-time Polymerase Chain Reaction, Control, Spectrophotometry

Journal: STAR Protocols

Article Title: Protocol for high-resolution 3D spatial transcriptomics using Open-ST

doi: 10.1016/j.xpro.2024.103521

Figure Lengend Snippet: Exonuclease I mix for flow cell processing

Article Snippet: Alkaline phosphatase calf intestinal (CIAP) enzyme , Promega , Cat#M1821.

Techniques: Concentration Assay

Journal: mSphere

Article Title: Chlamydomonas IC97, an intermediate chain of the flagellar dynein f/I1, is required for normal flagellar and cellular motility

doi: 10.1128/msphere.00558-24

Figure Lengend Snippet: The abundance of IDA-f/I1-HCs/LCs is relatively normal, but the phosphorylation of IC138 is apparently increased in the ic97 mutant. ( A ) Urea-gel PAGE of dynein HCs from axonemes of the wild type, the ic97 mutant, the rescued strain, and ida1 . The ic97 mutant has the HCα of IDA f/I1 (red circle), which is missing in ida1 (white circle). The other HC (HCβ) of IDA f/I1 overlaps with the HCγ of ODA and cannot be observed in this gel. The relative position of the lanes has been adjusted for presentation, and all lanes are from the same gel. ( B ) SDS-PAGE patterns of LCs (~10 kDa) of purified IDA f/I1 from axonemes of oda1 , the ic97 mutant, and the rescued strain using Uno-Q ion-exchange chromatography. For this analysis, we used oda1 instead of the wild type to facilitate the separation of IDA f/I1. There is no major difference in LC (~10 kDa) composition between these strains. All lanes are from different gels, and the development time of silver staining, and the contrast and/or brightness of the images were adjusted to clearly observe the LCs on each gel. ( C ) Western blotting of the wild type, the ic97 mutant, and mia2 axonemes using the IC138 antibody. Axonemes were treated with or without calf intestinal alkaline phosphatase (CIAP) for ~1 hour prior to analysis. The mia2 mutant has been shown to have hyperphosphorylated IC138 in flagella ( , ). The phosphorylation state of IC138 in the ic97 mutant appears to be intermediate between the wild type and mia2 .

Article Snippet: Calf-intestinal alkaline phosphatase (CIAP) treatment of axonemes ( , ) was performed by adding excess amounts (>1 unit per 1 µg protein) of CIAP (Promega) to axonemes in HMDENa buffer (30 mM HEPES, 5 mM MgSO 4 , 1 mM DTT, 1 mM EGTA, 50 mM NaCl [pH7.4]) and incubating the axonemes with CIAP for ~1 hour at room temperature.

Techniques: Mutagenesis, SDS Page, Purification, Ion Exchange Chromatography, Silver Staining, Western Blot