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. 2023 Jul 5;14:3958. doi: 10.1038/s41467-023-39761-2

Fig. 4. Mutation of LRRC4/SH3PXD2A/SLC26A8 increases vasoactive neuropeptides in human neural cells.

Fig. 4

a–c The relative mRNA expression levels of PACAP, VIP, NPY, CGRPβ, TAC1, CALR, and SST in human neural cells transfected respectively with LRRC4 (a), SH3PXD2A (b), and SLC26A8 (c) mutant/wild-type (WT)/Control vector plasmids (n = 4 biologically independent experiments). a PACAP: Mutant vs WT, P = 0.0009; WT vs Vector, P = 0.9992, VIP: Mutant vs WT, P = 0.0094; WT vs Vector, P = 0.9961. b PACAP: Mutant vs WT, P = 0.0324; WT vs Vector, P > 0.9999, NPY: Mutant vs WT, P = 0.0064; WT vs Vector, P = 0.9874, TAC1: Mutant vs WT, P = 0.0005; WT vs Vector, P = 0.9795. c PACAP: Mutant vs WT, P = 0.0004; WT vs Vector, P > 0.9999, VIP: Mutant vs WT, P = 0.0086; WT vs Vector, P = 0.9417, CGRPβ: Mutant vs WT, P < 0.0001; WT vs Vector, P = 0.9866, CALR: Mutant vs WT, P = 0.0006; WT vs Vector, P = 0.9996, SST: Mutant vs WT, P = 0.0004; WT vs Vector, P = 0.5298. d Immunostaining of PACAP in human neural cells transfected respectively with LRRC4, SH3PXD2A, and SLC26A8 mutant/WT/control vector plasmids. Right panels, the quantification of mean fluorescent intensity for PACAP in the corresponding groups. n = 42–71 cells from three independent experiments. LRRC4: Mutant vs WT, P < 0.0001; WT vs Vector, P = 0.9976, SH3PXD2A: Mutant vs WT, P < 0.0001; WT vs Vector, P = 0.5517, SLC26A8: Mutant vs WT, P < 0.0001; WT vs Vector, P = 0.0612. e Co-immunostaining of PACAP and PGP9.5 on skin sections from rosacea patients (rosacea, n = 6) and healthy individuals (HS, n = 5). Higher-magnified images of yellow boxed areas are shown below the lower-magnified images for each group. Scale bar, 50 μm. Yellow arrowheads indicate PGP9.5 positive neuron axon with strong co-immunostaining signals of PACAP; White arrowheads indicate PGP9.5 positive neuron axon with low or no immunostaining signals of PACAP. f Quantification of mean fluorescent intensity for PACAP in PGP9.5 positive neuron fibers (n = 25 for HS; n = 35 for rosacea). P < 0.0001. DAPI staining (blue) indicates nuclear localization. Scale bar, 50 μm. All results are representative of at least three independent experiments. Data represent the mean ± SEM. *P < 0.05, **P < 0.01. ns indicates no significance. One-way ANOVA with Bonferroni’s post hoc test (a–d) or two-tailed unpaired Student’s t test (f) was used.