Journal: Molecular Medicine
Article Title: Lhx6 deficiency causes human embryonic palatal mesenchymal cell mitophagy dysfunction in cleft palate
doi: 10.1186/s10020-024-00960-2
Figure Lengend Snippet: Abnormal mitophagy inhibited the proliferation and migration of HEPM cells by regulating MAPK signaling pathway. ( A ) A volcanic plot representing significantly altered gene numbers in the shCtrl vs. shLhx6 group. ( B ) A Venn diagram showing the distribution of genes in the shCtrl vs. shLhx6 group. ( C ) Enriched KEGG pathway analysis between the shCtrl and shLhx6 group. ( D ) Column-clustered heat map of MAPK signaling pathway-related genes that are differentially expressed between the two groups. The color intensity represents the relative mRNA level; red and blue indicate high and low expression, respectively. ( E , F , G ) Representative western blots and densitometric analyses showing the p38 and ERK1/2 MAPK signal pathway proteins in each group. ( H ) qPCR analysis to confirm the efficiency of PINK1 knockdown. ( I ) Representative western blots and densitometric analyses showing the p38 and ERK1/2 MAPK signal pathway proteins in each group. Data in the bar diagrams are represented as mean ± SD. N = at least 3 biological replicates. Circles correspond to each tested individual. NS P > 0.05, * P < 0.05, ** P < 0.01, *** P < 0.001
Article Snippet: After blocking the membranes with 5% bovine serum albumin (Biofroxx, Germany) at room temperature for 1 h, the membranes were incubated at 4°C overnight with primary antibodies: polyclonal rabbit anti-Lhx6 (1:500, Bioss, China), monoclonal rabbit anti-LC3 (1:1000, CST, USA), polyclonal rabbit anti-Beclin1 (1:500, Boster, China), polyclonal rabbit anti-p62 (1:500, Servicebio, China), monoclonal rabbit anti-ATG5 (1:1000, CST, USA), polyclonal rabbit anti-PINK1 (1:500, Affinity Biosciences, China), polyclonal rabbit anti-Parkin (1:500, Affinity Biosciences, China), polyclonal rabbit anti-TOMM20 (1:500, Affinity Biosciences, China), monoclonal rabbit anti-Phospho-p38 MAPK (1:1000, CST, USA), monoclonal rabbit anti-p38 MAPK (1:1000, CST, USA), monoclonal rabbit anti-Phospho-p44/42 MAPK (ERK1/2) (1:1000, CST, USA), monoclonal rabbit anti-p44/42 MAPK (ERK1/2) (1:1000, CST, USA), polyclonal rabbit anti-β-actin (1:1000, Affinity Biosciences, China).
Techniques: Migration, Expressing, Western Blot, Knockdown