Skip to main content
. 2018 Nov 20;5:99. doi: 10.3389/fmolb.2018.00099

Figure 3.

Figure 3

CaMKK2 is not directly associated with TF and loss of CaMKK2 affected TF associated protein complexes. (A) Immunoblot showing expression of CaMKK2 in the wild-type and three CRISPR/Cas9 based CaMKK2 KO HEK293 clonal cell lines selected independently (Top panel). Bottom panel: Flag-TF and GAPDH expression in HEK293 cells. (B) Immunoblots showing charged fractions of Flag-TF in the wild-type and CaMKK2 KO HEK293 cells. The red arrow represents P-TF. Blue square represents HMW TF. (C) Scatter plot showing the relative amount of P-TF. N = 4 from 4 independent experiments. The p-value by t-test (unpaired). (D) Immunoblots showing the presence of the TF and absence of the CaMKK2 in the Flag-TF immunoprecipitated fraction. The same protein fraction was also immunoblotted with the pan-P-ser specific antibody. (E) Immunoblots showing the relative shift of crosslinked CaMKK2 and TF in Flag-TF expressed wild-type and CaMKK2 KO HEK293 cells. The red squares represent the relative shift of the crosslinked proteins. Black Square represents native non-crosslinked form, shifted bands are numbered in Arabic numerals. (F,G) Immunoblots showing TF and CaMKK2 associated MPCs in mouse wild-type and CaMKK2 KO DRG tissues and Flag-TF expressed wild-type and CaMKK2 KO HEK293 cells. Dotted vertical lines indicate vertical alignment of the co-migrated protein complexes. Colored circles indicate MPCs. ns, Non-specific. The Coomassie stained gel strip on the top panel showing native page molecular weight ladder.