Total RNAs were collected from wing discs of MS1096-Gal4 (black bar), MS1096-Gal4 > lola RNAi-1 (red bar), MS1096-Gal4 > lola RNAi-2 (blue bar), and MS1096-Gal4 > lola RNAi-3 (gray bar). Note a significant decrease in the mRNA levels when lola RNAi is expressed (**p
(A) Relative enrichment of Lola on binding and non-binding (genebody, gb) regions compared to IgG in control (Renilla dsRNA treated, gray and black bars) and Lola knockdown cells (lola dsRNA treated, pink and red bars) were analyzed by ChIP and real-time PCR. Note a significant increase of Lola enrichment in regions near pdm3, ec, Dref and SkpA (ns p>0.05, *p
The efficiencies of RNAi lines for wts, lola, Dref, and SkpA have been verified by qPCR. As shown in Author response image 2, the expression levels of these genes were efficiently repressed. We have added the references of several RNAi transgenes in the Key Resources Table in the Materials and methods section.
We thank the reviewer for the suggestion. lola overexpression in ISCs represses DSS-induced proliferation as shown by Author response image 3. This piece of evidence indicates that Lola is functionally as relevant as Yki in ISCs. However, despite the repression, the fact that lola overexpression artificially increases Lola protein levels might have led to repression in any format.
The outer membrane-directed lipoproteins are released from the inner membrane of Escherichia coli as a complex with LolA, a periplasmic chaperone. The LolA-dependent release of lipoproteins is critical for lipoprotein sorting as it depends on the outer membrane-specific sorting signal. To clarify molecular events involved in the LolA-dependent lipoprotein release, we attempted to establish an in vitro assay system. The major outer membrane lipoprotein (Lpp) was found to lose its release competence soon after being processed to mature Lpp in the inner membrane and therefore could not be used as a substrate for an in vitro system. An Lpp derivative, L10P, was constructed and found to retain the release competence long after its maturation. L10P was synthesized and radiolabeled in spheroplasts in the absence of LolA; therefore, it remained anchored to the inner membrane of spheroplasts. Right-side out membrane vesicles containing L10P were then prepared and used to examine the release of L10P. In addition to LolA, L10P release absolutely required nucleoside triphosphate (NTP). A non-hydrolyzable NTP analogue strongly inhibited the NTP-dependent release. The outer membrane-specific sorting signal was essential for the in vitro release of L10P. Furthermore, L10P released in vitro was specifically incorporated into the outer membrane. These results indicate that the in vitro release of L10P represents an in vivo reaction and requires energy.
Somewhat shy, Lola is a 4-year-old pit bull-greyhound mix. Volunteers and staff are spending lots of time with her, and their efforts are paying off. This black beauty is really starting to come out of her shell. She is housebroken and good on leash. She should be the only pet with a mature family. To adopt Lola, request A4263843. The Agoura Shelter is at 29525 Agoura Road. Occasionally pets have already been adopted. To check availability, visit www.animalcare.lacounty.gov or call 818-991-0071.
Ely is a mature boy who has lots of life and love to give. He was found wandering the streets and taken to PAWS Place Adoption Center in Anna after weeks of searching for his home. He is a lover who is always happy and ready to play. He is about 8 or 9 years old and weighs about 8 pounds. 041b061a72