FIGURE

Fig. 6

ID
ZDB-FIG-221022-6
Publication
Zhao et al., 2022 - Brain milieu induces early microglial maturation through the BAX-Notch axis
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Fig. 6

The CaMKII-CREB pathway regulates dlb expression and microglial maturation.

a Immunofluorescence of p-CaMKII (magenta), H-G (green) and c-D (red) in the transverse sections of 3 dpf WT midbrain. p-CaMKII co-stained with GFP+ (white arrowhead) but not DsRed+ (white arrow) signals. b Immunofluorescence of p-CREB (magenta), H-G (green) and c-D (red) in the transverse sections of 3 dpf WT midbrain. p-CREB merged with GFP+ (white arrowhead) but not DsRed+ (white arrow) signals. c WB of p-CaMKII, CaMKII (pan), p-CREB, CREB (pan), and Tubulin in the brain cells from 3 dpf siblings and baxcq55 mutants. The experiments were repeated three times and the results were similar. d WISH of dlb in 3 dpf midbrains after treated with DMSO or PF-CBP1 HCl. e The number of c-D+ and a-G+c-D+ signals in 3 dpf midbrains after DMSO and PF-CBP1 HCl treatment. Each dot denotes one fish (n = 6 in each group). f Representative images of dlb and apoeb WISH, and NR staining in the baxcq55 midbrains after supplying NBT:creb1a. g Top panel: the predicted schematic structure of dlb promoter with two potential CREB binding sites. Bottom panel: the gel results of ChIP-PCR. p-CREB binding sites 1 and 2 were amplified by the dlb-F1/R1 and dlb-F2/R2 primers, respectively. dlb-NF/NR primers were used as the negative control. h Quantification of fold alterations in the luciferase activities after transfected with different vectors. Data were pooled from three independent experiments. Each dot represents an independent experiment. i A diagram of the CaMKII-CREB axis in controlling dlb expression, created with BioRender.com. MU, mutation. Numbers in the right corners in (d) and (f) indicate the counts of embryos with a typical appearance (first number) in the total examined fishes (last number). Error bars, mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns no significant, Unpaired two-tailed Student’s t test. Source data are provided as a Source Data file.

Expression Data

Expression Detail
Antibody Labeling
Phenotype Data

Phenotype Detail
Acknowledgments
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